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Sample records for vitro skin penetration

  1. Penetration of the laser light into the skin in vitro.

    Science.gov (United States)

    Kolárová, H; Ditrichová, D; Wagner, J

    1999-01-01

    Knowledge of the optical parameters of the skin is important for all kinds of phototherapy. We analyzed penetration of laser light and proved different optical properties of in vitro specimens of normal skin and granular tissue from skin ulcers. An He-Ne laser (lambda = 632.8 nm, output 50 mW) and a semiconductor laser (lambda = 675 nm, output 21 mW) were used. The distribution of laser radiation was detected by a CCD camera and evaluated by the image analysis software DIPS. Transmittance in granular tissue was about 2.5 times higher than that in normal skin. In the thickest skin sample (2 cm), approximately 0.3% of He-Ne laser and 2.1% of semiconductor laser light penetrated. The results demonstrate the percentage of incident light penetrating the individual skin layers in different localizations on the skin surface, which is a decisive factor for the selection of the radiation dose.

  2. Skin penetration enhancement by a microneedle device (Dermaroller) in vitro

    DEFF Research Database (Denmark)

    Badran, M M; Kuntsche, Judith; Fahr, A

    2009-01-01

    This study focused on the in vitro evaluation of skin perforation using a new microneedle device (Dermaroller) with different needle lengths (150, 500 and 1500 microm). The influence of the microneedle treatment on the morphology of the skin surface (studied by light and scanning electron...... microscopy), on the transepidermal water loss (TEWL) and on the penetration and permeation of hydrophilic model drugs was investigated using excised human full-thickness skin. Furthermore, invasomes - highly flexible phospholipid vesicles containing terpenes and ethanol as penetration enhancer - were...... compared with an aqueous solution. Elevated TEWL values were measured after Dermaroller treatment compared to untreated human skin with a gradual increase of the TEWL over the first hour whereas afterwards the TEWL values decreased probably caused by a reduction of the pore size with time. Skin perforation...

  3. Evaluation of critical parameters for in vitro skin permeation and penetration studies using animal skin models.

    Science.gov (United States)

    Praça, Fabíola Silva Garcia; Medina, Wanessa Silva Garcia; Eloy, Josimar O; Petrilli, Raquel; Campos, Patrícia Mazureki; Ascenso, Andreia; Bentley, Maria Vitória L B

    2017-09-23

    In vitro skin permeation/penetration studies may be affected by many sources of variation. Herein, we aimed to investigate the major critical procedures of in vitro skin delivery studies. These experiments were performed with model drugs according to official guidelines. The influence of skin source on penetration studies was studied as well as the use of a cryopreservation agent on skin freezing evaluated by transepidermal water loss, electrical resistance, permeation/penetration profiles and histological changes of the skin. The best condition for tape stripping procedure was validated through the evaluation of the distribution of corneocytes, mass of stratum corneum (SC) removed and amount of protein removed using finger pressure, a 2kg weight and a roller. The interchangeability of the tape stripping procedures followed by the epidermis and dermis homogenate and the micrometric horizontal cryostat skin sectioning methods were also investigated, besides the effect of different formulations. Noteworthy, different skin sources were able to ensure reliable interchangeability for in vitro permeation studies. Furthermore, an increased penetration was obtained for stored frozen skin compared to fresh skin, even with the addition of a cryoprotectant agent. The best method for tape stripping was the finger pressure followed by the addition of a propylene glycol solvent leading to better SC removal. Finally, no significant difference was found in skin penetration studies performed by different methods suggesting their possible interchangeability. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Storage conditions of skin affect tissue structure and in vitro percutaneus penetration

    DEFF Research Database (Denmark)

    Nielsen, Jesper Bo; Bagatolli, Luis

    2017-01-01

    skin at -20oC causes structural changes in the upper Stratum Corneum observable with image techniques such as multiphoton excitation fluorescence microscopy. The presently available literature does, however, not support that the observed structural damage to the integrity is sufficient to cause......For logistic and practical reasons it is difficult to perform in vitro studies on percutaneous penetration on fresh human skin obtained directly from surgery. Skin samples are therefore often kept frozen until use. The present chapter present the available literature on the topic. Storage of human...... a general and significantly increased in vitro percutaneous penetration across human skin stored at -20oC. Use of skin stored at -20oC for in vitro studies on percutaneous penetration therefore seems acceptable as long as the barrier integrity is documented. Storage of human skin at -80oC causes significant...

  5. Storage Conditions of Skin Affect Tissue Structure and Subsequent in vitro Percutaneous Penetration

    DEFF Research Database (Denmark)

    Nielsen, J B; Plasencia, I; Sørensen, J A

    2011-01-01

    fluorescence microscopy) and in vitro percutaneous penetration of caffeine under four different storage conditions using skin samples from the same donors: fresh skin, skin kept at -20°C for 3 weeks (with or without the use of polyethylene glycol) and at -80°C. Our results show a correlation between increasing...... permeation of caffeine and tissue structural damage caused by the storage conditions, most so after skin storage at -80°C. The presented approach, which combines imaging techniques with studies on percutaneous penetration, enables the link between tissue damage at selected depths and penetration...

  6. Increased in vivo skin penetration of quantum dots with UVR and in vitro quantum dot cytotoxicity

    Science.gov (United States)

    Mortensen, Luke; Zheng, Hong; Faulknor, Renea; De Benedetto, Anna; Beck, Lisa; DeLouise, Lisa A.

    2009-02-01

    The growing presence of quantum dots (QD) in a variety of biological, medical, and electronics applications means an increased risk of human exposure in manufacturing, research, and consumer use. However, very few studies have investigated the susceptibility of skin to penetration of QD - the most common exposure route- and the results of those that exist are conflicting. This suggests that a technique allowing determination of skin barrier status and prediction of skin permeability to QD would be of crucial interest as recent findings have provided evidence of in vitro cytotoxicity and long-term in vivo retention in the body for most QD surface chemistries. Our research focuses on barrier status of the skin (intact and with ultraviolet radiation induced barrier defect) and its impact on QD skin penetration. These model studies are particularly relevant to the common application condition of NP containing sunscreen and SPF cosmetics to UV exposed skin. Herein we present our initial efforts to develop an in vivo model of nanoparticle skin penetration using the SKH-1 hairless mouse with transepidermal water loss (TEWL) to evaluate skin barrier status and determine its ability to predict QD penetration. Our results show that ultraviolet radiation increases both TEWL and skin penetration of QD. Additionally, we demonstrate cytotoxic potential of QD to skin cells using a metastatic melanoma cell line. Our research suggests future work in specific targeting of nanoparticles, to prevent or enhance penetration. This knowledge will be used to develop powerful therapeutic agents, decreased penetration cosmetic nanoparticles, and precise skin cancer imaging modalities.

  7. Enhancement of percutaneous penetration of aniline and o-toluidine in vitro using skin barrier creams.

    Science.gov (United States)

    Korinth, Gintautas; Lüersen, Lars; Schaller, Karl Heinz; Angerer, Jürgen; Drexler, Hans

    2008-04-01

    Aniline (ANI) and the human carcinogen o-toluidine (OT) are released at the workplace during the production and processing of rubber. Recently, we showed in rubber industry workers that a frequent use of skin barrier creams (SBC) increased the internal exposure of ANI and OT. In the present study, diffusion cells were used to investigate the effects of two SBC and one skin care cream (SCC) on percutaneous penetration of neat ANI and OT as well as of OT from a mixture with a workplace specific lubricant. The experiments were carried out with untreated and with skin creams treated human skin. A considerable percutaneous penetration enhancement of test compounds was observed for treated skin compared with untreated skin; the highest enhancement (mean factors 6.2-12.3) was found for SBC (based on oil in water emulsion) treated skin. The lowest penetration enhancement showed SCC treated skin (mean factors 4.2-9.7). The in vitro data support our findings in workers that the percutaneous absorption of aromatic amines significantly increases in presence of skin creams. The efficacy of skin creams to protect the percutaneous penetration of aromatic amines is not confirmed by our own experiments.

  8. In vitro skin penetration of acetyl hexapeptide-8 from a cosmetic formulation.

    Science.gov (United States)

    Kraeling, Margaret E K; Zhou, Wanlong; Wang, Perry; Ogunsola, Oluwatosin A

    2015-03-01

    There is a concern that peptides in cosmetic creams marketed as anti-aging/anti-wrinkle may penetrate into the deep layers of the skin and potentially stimulate biological activity. Claims for one cosmetic peptide, acetyl hexapeptide-8 (Ac-EEMQRR-amide), suggest interference with neuromuscular signaling as its anti-wrinkle mechanism of action. Therefore, the skin penetration of commercially available Ac-EEMQRR-amide from a cosmetic formulation (oil-in-water (O/W) emulsion) was determined in hairless guinea pig (HGP) and human cadaver skin assembled into in vitro diffusion cells. An O/W emulsion containing 10% Ac-EEMQRR-amide was applied to skin at a dose of 2 mg/cm(2). After a 24-h exposure, the skin surface was washed to remove unabsorbed peptide. Skin disks were tape stripped to determine the amount of peptide in the stratum corneum. Removal of the stratum corneum layers was verified by confocal microscopy. The epidermis was heat separated from the dermis and each skin fraction was homogenized. Skin penetration of Ac-EEMQRR-amide was measured in skin layers by hydrophilic interaction liquid chromatography with tandem mass spectrometry using electrospray ionization (ESI) in the positive mode. Stable isotopically labeled hexapeptides were used as internal standards for the quantitation of native hexapeptides to correct for matrix effects associated with ESI. The results (percent of applied dose) showed that the majority of the Ac-EEMQRR-amide was washed from the surface of both HGP and human skin. Ac-EEMQRR-amide that penetrated skin remained mostly in the stratum corneum of HGP (0.54%) and human (0.22%) with the peptide levels decreasing as each layer was removed by tape stripping. Total Ac-EEMQRR-amide found in the epidermis of HGP and human skin was similar at 0.01%. No peptide was detected in the dermis or buffer collected underneath the skin for both human and HGP. There was no hexapeptide metabolite (H2N-EEMQRR-amide) detected in any layers of HGP skin, human

  9. In vitro study of ethosome penetration in human skin and hypertrophic scar tissue.

    Science.gov (United States)

    Zhang, Zhen; Wo, Yan; Zhang, Yixin; Wang, Danru; He, Rong; Chen, Huijin; Cui, Daxiang

    2012-08-01

    The purpose of this study is to characterize a novel transdermal delivery carrier, ethosomes containing 5-fluorouracil. The delivery of drugs from ethosomes in human hypertrophic scar (HS) and the mechanisms of action of ethosomes in human HS were investigated. Percutaneous ethosome permeation was evaluated in vitro in human HS and skin using a Franz's cell. The amount of 5-fluorouracil that permeated HS and skin after 24 hours was most abundant in ethosomes via HS (E-Scar), followed by hydroethanolic solution via HS (H-Scar), ethosomes via skin (E-Skin), and hydroethanolic solution via skin (H-Skin). The penetration of ethosomes in HS and skin was analyzed by ethosomes fluorescently labeled with rhodamine 6GO using confocal laser scanning microscopy. The fluorescence intensity after application for 24 hours was highest in E-Scar, followed by E-Skin, H-Scar, and H-Skin, which indicates the penetration of ethosomes in HS was greatest. In conclusion, we consider that ethosomes are a highly efficient carrier in HS. Copyright © 2012 Elsevier Inc. All rights reserved.

  10. [Physicochemical properties and skin penetration in vitro of total alkaloids of Sophora flavescens nanoemulsion].

    Science.gov (United States)

    Feng, Ai-Ling; Wang, Ying-Zi; Zhang, Sheng-Hai; Sun, Xiu-Yu; Duan, Fei-Peng; Li, Cai-Xia

    2013-08-01

    The research aimed at investigating the physicochemical properties, stability and skin penetration in vitro of total alkaloids of Sophora flavescens nanoemulsion. Prepare total alkaloids of S. flavescens nanoemulsion and detect the determination of matrine and oxymatrine in the nanoemulsion using HPLC method. Transmission electron microscopy and laser particle size analyzer were utilized to detect the shape and size of the nanoemulsion respectively. And also the stability of nanoemulsion was studied under the conditions of low temperature (4 degrees C), normal temperature (25 degrees C) and high temperature (60 degrees C). Franz diffusion cell was used to research the transdermal absorption of nanoemulsion in vitro. The results found that the nanoemulsion we prepared presented appearance of rounded, uniform; its average diameter was (15.55 +/- 2.24) nm, and particle size distribution value was 0. 161; the appearance, diameter and percentage determination of total alkaloids of S. flavescens had no variations after 15 d under 4, 25, 60 degrees C respectively. The steady-state permeation rate was 4.564 1 microg x cm(-2) x h(-1), 24 h cumulative amount of penetration was 110.7 microg x cm(-2), which was 1.86 fold of 24 h cumulative amount of aqueous solution (59.41 microg x cm(-2)). All the results demonstrated total alkaloids of S. flavescens nanoemulsion had good permeability, and could provide a new preparation for its clinical application.

  11. In vitro investigation of the follicular penetration of porcine ear skin using a nanoparticle-emulsion containing the antiseptic polihexanide In vitro investigation of the follicular penetration of porcine ear skin

    Science.gov (United States)

    Ulmer, M.; Patzelt, A.; Vergou, T.; Lademann, J.; Richter, H.; Kramer, A.; Müller, G.; Sterry, W.; Lange-Asschenfeldt, B.

    2012-05-01

    Earlier investigations regarding the distribution of the bacterial flora on the human skin demonstrate that the hair follicle acts as a bacterial reservoir, providing a quick source for secondary recontamination. These findings highlight the importance of the hair follicle as a target for modern antiseptics. In the present study, we have assessed the follicular penetration of a curcumin-labeled particle-associated antiseptic into porcine skin by laser scanning microscopy. Therefore, the follicular penetration depth of the curcumin-labeled particle-associated antiseptic was compared to the follicular penetration depth of curcumin-labeled particles without antiseptic. The investigation was performed in vitro using porcine skin biopsies. By superposition of the images acquired in the transmission and the fluorescent modus, it was possible to visualize the distribution of the fluorescent dye inside the hair follicles. Quantitative and qualitative results showed that both dispersions penetrated efficiently into the hair follicles. The average penetration depth of the particles with attached antiseptic polihexanide was significantly higher than that of particles without the attached antiseptic. Also, whilst very little sample preparation was needed, laser scanning microscopy was found to be an efficient tool to visualize the skin relief and in particular the hair follicle shaft and localize fluorescent markers within the skin tissue and hair follicles.

  12. Comparative study of liposomes, transfersomes, ethosomes and cubosomes for transcutaneous immunisation: characterisation and in vitro skin penetration.

    Science.gov (United States)

    Rattanapak, Teerawan; Young, Katie; Rades, Thomas; Hook, Sarah

    2012-11-01

    Lipid colloidal vaccines, including liposomes, transfersomes, ethosomes and cubosomes, were formulated, characterised and investigated for their ability to enhance penetration of a peptide vaccine through stillborn piglet skin in vitro. Liposomes and transfersomes were formulated using a film-hydration method, ethosomes using a modified reverse phase method and cubosomes using a lipid precursor method. The size, zeta potential, peptide loading and interfacial behaviour of the formulations were characterised. Skin penetration studies were performed using Franz diffusion cells with piglet skin as the membrane. The localization of peptide in the skin was examined using confocal laser scanning microscopy. The various formulations contained negatively charged particles of similar size (range: 134-200 nm). Addition of the saponin adjuvant Quil A to the formulations destabilised the monolayers and reduced peptide loading. Cubosomes and ethosomes showed superior skin retention compared with the other systems. Confocal laser scanning microscopy showed greater peptide penetration and accumulation in the skin treated with cubosomes and ethosomes. With the other systems peptide was only located in the vicinity of the hair follicles and within the hair shaft. We conclude from the in-vitro studies that cubosomes and ethosomes are promising lipid carriers for transcutaneous immunisation. © 2012 The Authors. JPP © 2012 Royal Pharmaceutical Society.

  13. Influence of cosmetics vehicles on 4-methylbenzylidene-camphor's skin penetration, in vitro

    Directory of Open Access Journals (Sweden)

    Clarice Scliar Sasson

    2009-04-01

    Full Text Available The aim of this work was to compare the skin penetration of 4-methylbenzylidene-camphor (4-MBC in two vehicles, an oil-in-water (O/W emulsion and an alcoholic gel. The penetration of this UVB filter through these vehicles was determined in vitro (Franz cells using pig ear skin. The 4-MBC permeated through the skin both with the emulsion o/w as the alcoholic gel. However, with the alcoholic gel, 5 h after application of the product, the 4-MBC was found in the receptor fluid, while with the emulsion o/w, 24 h after it was detected. In both vehicles, the 4-MBC was present in the viable epidermis, dermis but most part of it, was found in the stratum corneum, being more remarkable for alcoholic gel. The right choice of the vehicle could decrease the potential toxicological risk and increase the efficacy of sunscreens products.O objetivo deste estudo foi avaliar a influência de diferentes veículos cosméticos na permeação cutânea da 4-metilbenzilideno cânfora através de experimento in vitro utilizando células de difusão de Franz. Os veículos avaliados foram um gel alcoólico e uma emulsão não iônica O/A. Durante o experimento, foi doseada a presença da 4-MBC no fluido receptor em µg/cm², desde o tempo 0 h até 24 h após a aplicação dos veículos, bem como sua concentração na superfície cutânea, estrato córneo e [epiderme sem estrato córneo + derme]. No estudo realizado com o gel alcoólico, a partir de 5 h após o início do experimento, já foi detectada a presença da 4-MBC no fluido receptor, ao passo que, utilizando a emulsão O/A, a 4-MBC foi detectada apenas após 24 h. Em ambos os veículos a 4-MBC estava presente na [epiderme sem estrato córneo + derme] não havendo diferença estatística entre eles. A maior concentração de 4-MBC foi detectada na superfície cutânea, estando em maior quantidade quando utilizada a emulsão O/A como veículo. Já no estrato córneo, a concentração de 4-MBC foi maior na utiliza

  14. In vitro skin penetration of clobetasol from lipid nanoparticles: drug extraction and quantitation in different skin layers

    Directory of Open Access Journals (Sweden)

    Luís Antônio Dantas Silva

    2012-12-01

    Full Text Available Clobetasol propionate (CP is a potent topical corticosteroid that causes several cutaneous and systemic side effects. In the present work, CP was encapsulated in nanostructured lipid carriers (NLCs to increase drug retention in the outer skin layers and improve the safety of topical therapy. NLCs were prepared using a microemulsion technique with a mixture of lecithin, taurodeoxycholate, stearic acid, and oleic acid. In vitro penetration studies were performed in a modified Franz-type diffusion cell, and porcine ears were used as a model of human skin. A simple and sensitive liquid chromatographic method was developed and validated for clobetasol determination in different skin layers. NLCs presented uniform size distribution, high zeta potentialand entrapment efficiency values (> 98%. The analytical procedure was validated according to FDA guidelines. Clobetasol recoveries from skin samples were higher than 85%, with no interference of skin components and NLC ingredients. In experiments, after 6 h, a higher drug accumulation in the stratum corneum arising from NLCs compared to aqueous CP solution was observed. Thus, the NLCs demonstrated high potential for targeting CP to the skin and ensuring drug accumulation in the stratum corneum.Proprionato de clobetasol (CP é um potente corticóide tópico, que apresenta vários efeitos adversos cutâneos e sistêmicos. No presente trabalho, CP foi encapsulado em carreadores lipídicos nanoestruturados (NLCs visando aumentar a retenção do fármaco nas camadas superficiais da pele e a segurança da terapia tópica. NLCs foram preparados usando a técnica de diluição de microemulsão com mistura de lecitina, taurodesoxicolato, ácido esteárico e ácido oléico. Estudos de penetração in vitro foram realizados em células de difusão de Franz modificadas usando pele de orelha de porco como modelo de pele humana. Um método simples e sensível de cromatografia líquida foi desenvolvido e validado para

  15. Fluorometric quantification of protoporphyrin IX in biological skin samples from in vitro penetration/permeation studies

    Directory of Open Access Journals (Sweden)

    Fábia Cristina Rossetti

    2010-12-01

    Full Text Available A fluorometric analytical method was developed for quantification of protoporphyrin IX (PpIX in skin samples and receptor phase solution after in vitro cutaneous penetration/permeation studies. Analytical conditions used were: excitation and emission wavelengths: 400 nm and 632 nm; bandwidth: 0.5 nm; excitation and emission slits: 10/10. PpIX was recovered from two different layers of skin, the stratum corneum (SC and the epidermis plus dermis ([E+D], by vortex homogenization, probe and bath sonication, using DMSO as an extraction solvent. The detection and quantification limits were 0.002 and 0.005 μg/mL, respectively. The assay was linear from 0.005 - 0.5 μg/mL. The within-day and between-day assay precision and accuracy in DMSO and receptor phase solution were each studied at the two concentration levels 0.04 and 0.2 μg/mL, and 0.01 and 0.08 μg/mL, respectively. The coefficients of variation and deviation from the theoretical values were lower than 5%. The skin recovery of PpIX from SC and [E+D] layers using two different concentrations (0.5 and 1.0 μg/mL were all above 90.0%. The method described has potential application to in vitro penetration/permeation studies of PpIX using porcine skin as a biological membrane model.Um método analítico por espectrofluorimetria foi desenvolvido para quantificar a protoporfirina IX (Pp IX em amostras de pele e fase receptora após a realização de testes in vitro de penetração/permeação cutâneas. As condições analíticas utilizadas foram: comprimentos de onda de excitação e emissão: 400 nm e 632 nm; largura de banda: 0,5 nm; fendas de excitação e emissão: 10/10. A PpIX foi extraída de amostras de estrato córneo (EC e da epiderme sem estrato córneo + derme ([E+D] através da agitação em vórtex e sonicação por haste e banho, utilizando-se o DMSO como solvente extrator. O limite de detecção e quantificação foram, respectivamente, de 0,002 e 0,005 μg/mL. O método mostrou

  16. Effect of elevating the skin temperature during topical ALA application on in vitro ALA penetration through mouse skin and in vivo PpIX production in human skin

    NARCIS (Netherlands)

    van den Akker, Johanna T. H. M.; Boot, Kristian; Vernon, David I.; Brown, Stanley B.; Groenendijk, Laurens; van Rhoon, Gerard C.; Sterenborg, Henricus J. C. M.

    2004-01-01

    An approach to induce increased protoporphyrin IX (PpIX) production in aminolevulinic acid (ALA)-based photodynamic therapy (PDT) of skin lesions is to elevate the skin temperature during topical ALA application. Increased skin temperature may increase the ( depth of) penetration of ALA into the

  17. Application of in vitro skin penetration measurements to confirm and refine the quantitative skin sensitization risk assessment of methylisothiazolinone.

    Science.gov (United States)

    Rothe, Helga; Ryan, Cindy A; Page, Leanne; Vinall, Joanne; Goebel, Carsten; Scheffler, Heike; Toner, Frank; Roper, Clive; Kern, Petra S

    2017-12-01

    Use of quantitative risk assessment (QRA) for assessing the skin sensitization potential of chemicals present in consumer products requires an understanding of hazard and product exposure. In the absence of data, consumer exposure is based on relevant habits and practices and assumes 100% skin uptake of the applied dose. To confirm and refine the exposure, a novel design for in vitro skin exposure measurements was conducted with the preservative, methylisothiazolinone (MI), in beauty care (BC) and household care (HHC) products using realistic consumer exposure conditions. A difference between measured exposure levels (MELs) for MI in leave-on versus rinse-off BC products, and lower MELs for MI in HHC rinse-off compared to BC products was demonstrated. For repeated product applications, the measured exposure was lower than estimations based on summation of applied amounts. Compared to rinse-off products, leave-on applications resulted in higher MELs, correlating with the higher incidences of allergic contact dermatitis associated with those product types. Lower MELs for MI in rinse-off products indicate a lower likelihood to induce skin sensitization, also after multiple daily applications. These in vitro skin exposure measurements indicate conservatism of default exposure estimates applied in skin sensitization QRA and might be helpful in future risk assessments. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Enhancement of the in vitro penetration of quercetin through pig skin by combined microneedles and lipid microparticles.

    Science.gov (United States)

    Paleco, Roberto; Vučen, Sonja R; Crean, Abina M; Moore, Anne; Scalia, Santo

    2014-09-10

    Silicon microneedle patches were investigated, alone or in combination with lipid microparticles (LMs), as a system to improve the in vitro skin penetration of the antioxidant flavonoid, quercetin. LMs loaded with quercetin were prepared by melt emulsification and sonication. The flavonoid content of LMs was 11.7±0.3% and their mean diameter and polydispersity index were 8.1 μm and 0.66, respectively. Emulsions containing quercetin, free or microencapsulated, were applied to untreated- or microneedle-treated pig skin mounted in Franz diffusion cells. The amount of flavonoid penetrated into the stratum corneum and viable epidermis were measured by HPLC, after validated tape-stripping and bead mill homogenization procedures, respectively. Compared to intact skin, a marked increase in quercetin levels permeated into the stratum corneum (from 1.19 ± 0.12 μg/cm(2) to 2.23 ± 0.54 μg/cm(2)) and viable epidermis (from 0.10 ± 0.01 μg/cm(2) to 0.56 ± 0.27 μg/cm(2)) was achieved when skin was treated with the flavonoid-loaded LMs in combination with microneedle arrays. Conversely, perforation of the cutaneous surface by microneedles did not produce any significant improvement in the skin penetration of non-encapsulated quercetin. The enhanced (5.5-fold) intra-epidermal delivery of quercetin attained by the LM/microneedle strategy described here, is particularly relevant since the main quercetin site of action is in the epidermis. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Penetration through the Skin Barrier

    DEFF Research Database (Denmark)

    Nielsen, Jesper Bo; Benfeldt, Eva; Holmgaard, Rikke

    2016-01-01

    and exogenous factors may affect barrier characteristics. The present chapter introduces the theory for barrier penetration (Fick's law), and describes and discusses different methods for measuring the kinetics of percutaneous penetration of chemicals, including in vitro methods (static and flow...

  20. Negligible penetration of incidental amounts of alpha-hydroxy acid from rinse-off personal care products in human skin using an in vitro static diffusion cell model.

    Science.gov (United States)

    Okuda, M; Donahue, D A; Kaufman, L E; Avalos, J; Simion, F A; Story, D C; Sakaguchi, H; Fautz, R; Fuchs, A

    2011-12-01

    Alpha-hydroxy acids (AHAs), primarily glycolic and lactic acids, are widely used in cosmetics to alleviate dyspigmentation, photodamage, and other aging skin conditions and as pH adjusters. Glycolic acid reportedly enhances skin damage after repeated ultraviolet light exposure, e.g., increased sunburn cell formation. This study assessed potential in vitro skin penetration of lactic acid and malic acid incorporated into rinse-off personal care products, compared with rinse-off and leave-on exposures to glycolic acid (10%, pH 3.5) in a reference lotion. Radiolabeled AHA-fortified shampoo, conditioner, and lotion were evenly applied as single doses to human epidermal membranes mounted in static diffusion cells (not occluded). Exposures were 1-3 min (rinse-off) or 24 h (leave-on). Epidermal penetration of malic acid and lactic acid from the rinse-off shampoo and conditioner, respectively, was negligible, with >99% removed by rinsing, a negligible portion remaining in the stratum corneum (≤0.15%), and even less penetrating into the viable epidermis (≤0.04%). Glycolic acid penetration from the leave-on reference lotion was 1.42 μg equiv./cm2/h, with total absorbable dose recovery (receptor fluid plus epidermis) of 2.51%, compared to 0.009%, 0.003%, and 0.04% for the rinse-off reference lotion, shampoo (malic acid), and conditioner (lactic acid) exposures, respectively. Dermal penetration of AHAs into human skin is pH-, concentration-, and time-dependent. Alpha-hydroxy acids in rinse-off shampoos and conditioners are almost entirely removed from the skin within minutes by rinsing (resulting in negligible epidermal penetration). This suggests that ultraviolet radiation-induced skin effects of AHA-containing rinse-off products are negligible. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. Vehicle effects on human stratum corneum absorption and skin penetration.

    Science.gov (United States)

    Zhang, Alissa; Jung, Eui-Chang; Zhu, Hanjiang; Zou, Ying; Hui, Xiaoying; Maibach, Howard

    2017-05-01

    This study evaluated the effects of three vehicles-ethanol (EtOH), isopropyl alcohol (IPA), and isopropyl myristate (IPM)-on stratum corneum (SC) absorption and diffusion of the [(14)C]-model compounds benzoic acid and butenafine hydrochloride to better understand the transport pathways of chemicals passing through and resident in SC. Following application of topical formulations to human dermatomed skin for 30 min, penetration flux was observed for 24 h post dosing, using an in vitro flow-through skin diffusion system. Skin absorption and penetration was compared to the chemical-SC (intact, delipidized, or SC lipid film) binding levels. A significant vehicle effect was observed for chemical skin penetration and SC absorption. IPA resulted in the greatest levels of intact SC/SC lipid absorption, skin penetration, and total skin absorption/penetration of benzoic acid, followed by IPM and EtOH, respectively. For intact SC absorption and total skin absorption/penetration of butenafine, the vehicle that demonstrated the highest level of sorption/penetration was EtOH, followed by IPA and IPM, respectively. The percent doses of butenafine that were absorbed in SC lipid film and penetrated through skin in 24 h were greatest for IPA, followed by EtOH and IPM, respectively. The vehicle effect was consistent between intact SC absorption and total chemical skin absorption and penetration, as well as SC lipid absorption and chemical penetration through skin, suggesting intercellular transport as a main pathway of skin penetration for model chemicals. These results suggest the potential to predict vehicle effects on skin permeability with simple SC absorption assays. As decontamination was applied 30 min after chemical exposure, significant vehicle effects on chemical SC partitioning and percutaneous penetration also suggest that skin decontamination efficiency is vehicle dependent, and an effective decontamination method should act on chemical solutes in the lipid domain.

  2. [Effects of Frankincense and Myrrh essential oil on transdermal absorption in vitro of Chuanxiong and penetration mechanism of skin blood flow].

    Science.gov (United States)

    Zhu, Xiao-Fang; Luo, Jing; Guan, Yong-Mei; Yu, Ya-Ting; Jin, Chen; Zhu, Wei-Feng; Liu, Hong-Ning

    2017-02-01

    The aim of this paper was to explore the effects of Frankincense and Myrrh essential oil on transdermal absorption in vitro of Chuanxiong, and to investigate the possible penetration mechanism of their essential oil from the perspective of skin blood perfusion changes. Transdermal tests were performed in vitro with excised mice skin by improved Franz diffusion cells. The cumulative penetration amounts of ferulic acid in Chuanxiong were determined by HPLC to investigate the effects of Frankincense and Myrrh essential oil on transdermal permeation properties of Chuanxiong. Simultaneously, the skin blood flows were determined by laser flow doppler. The results showed that the cumulative penetration amount of ferulic acid in Chuanxiong was (8.13±0.76) μg•cm⁻² in 24 h, and was (48.91±4.87), (57.80±2.86), (63.34±4.56), (54.17±4.40), (62.52±7.79) μg•cm⁻² respectively in Azone group, Frankincense essential oil group, Myrrh essential oil, frankincense and myrrh singly extracted essential oil mixture group, and frankincense and myrrh mixed extraction essential oil group. The enhancement ratios of each essential oil groups were 7.68, 8.26, 7.26, 8.28, which were slightly greater than 6.55 in Azone group. In addition, as compared with the conditions before treatment, there were significant differences and obvious increasing trend in blood flow of rats in Frankincense essential oil group, Myrrh essential oil group, frankincense and myrrh singly extracted essential oil mixture group, and frankincense and myrrh mixed extraction essential oil group when were dosed at 10, 20, 30, 10 min respectively, indicating that the skin blood flows were increased under the effects of Frankincense and Myrrh essential oil to a certain extent. Thus, Frankincense and Myrrh essential oil had certain effect on promoting permeability of Chuanxiong both before and after drug combination, and may promote the elimination of drugs from epidermis to dermal capillaries through increase of

  3. Enhanced chlorhexidine skin penetration with eucalyptus oil

    Directory of Open Access Journals (Sweden)

    Worthington Tony

    2010-09-01

    Full Text Available Abstract Background Chlorhexidine digluconate (CHG is a widely used skin antiseptic, however it poorly penetrates the skin, limiting its efficacy against microorganisms residing beneath the surface layers of skin. The aim of the current study was to improve the delivery of chlorhexidine digluconate (CHG when used as a skin antiseptic. Method Chlorhexidine was applied to the surface of donor skin and its penetration and retention under different conditions was evaluated. Skin penetration studies were performed on full-thickness donor human skin using a Franz diffusion cell system. Skin was exposed to 2% (w/v CHG in various concentrations of eucalyptus oil (EO and 70% (v/v isopropyl alcohol (IPA. The concentration of CHG (μg/mg of skin was determined to a skin depth of 1500 μm by high performance liquid chromatography (HPLC. Results The 2% (w/v CHG penetration into the lower layers of skin was significantly enhanced in the presence of EO. Ten percent (v/v EO in combination with 2% (w/v CHG in 70% (v/v IPA significantly increased the amount of CHG which penetrated into the skin within 2 min. Conclusion The delivery of CHG into the epidermis and dermis can be enhanced by combination with EO, which in turn may improve biocide contact with additional microorganisms present in the skin, thereby enhancing antisepsis.

  4. The isolated perfused human skin flap model: A missing link in skin penetration studies?

    Science.gov (United States)

    Ternullo, Selenia; de Weerd, Louis; Flaten, Gøril Eide; Holsæter, Ann Mari; Škalko-Basnet, Nataša

    2017-01-01

    Development of effective (trans)dermal drug delivery systems requires reliable skin models to evaluate skin drug penetration. The isolated perfused human skin flap remains metabolically active tissue for up to 6h during in vitro perfusion. We introduce the isolated perfused human skin flap as a close-to-in vivo skin penetration model. To validate the model's ability to evaluate skin drug penetration the solutions of a hydrophilic (calcein) and a lipophilic (rhodamine) fluorescence marker were applied. The skin flaps were perfused with modified Krebs-Henseleit buffer (pH7.4). Infrared technology was used to monitor perfusion and to select a well-perfused skin area for administration of the markers. Flap perfusion and physiological parameters were maintained constant during the 6h experiments and the amount of markers in the perfusate was determined. Calcein was detected in the perfusate, whereas rhodamine was not detectable. Confocal images of skin cross-sections shoved that calcein was uniformly distributed through the skin, whereas rhodamine accumulated in the stratum corneum. For comparison, the penetration of both markers was evaluated on ex vivo human skin, pig skin and cellophane membrane. The proposed perfused flap model enabled us to distinguish between the penetrations of the two markers and could be a promising close-to-in vivo tool in skin penetration studies and optimization of formulations destined for skin administration. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Setup for investigating gold nanoparticle penetration through reconstructed skin and comparison to published human skin data

    Science.gov (United States)

    Labouta, Hagar I.; Thude, Sibylle; Schneider, Marc

    2013-06-01

    Owing to the limited source of human skin (HS) and the ethical restrictions of using animals in experiments, in vitro skin equivalents are a possible alternative for conducting particle penetration experiments. The conditions for conducting penetration experiments with model particles, 15-nm gold nanoparticles (AuNP), through nonsealed skin equivalents are described for the first time. These conditions include experimental setup, sterility conditions, effective applied dose determination, skin sectioning, and skin integrity check. Penetration at different exposure times (two and 24 h) and after tissue fixation (fixed versus unfixed skin) are examined to establish a benchmark in comparison to HS in an attempt to get similar results to HS experiments presented earlier. Multiphoton microscopy is used to detect gold luminescence in skin sections. λex=800 nm is used for excitation of AuNP and skin samples, allowing us to determine a relative index for particle penetration. Despite the observed overpredictability of penetration into skin equivalents, they could serve as a first fast screen for testing the behavior of nanoparticles and extrapolate their penetration behavior into HS. Further investigations are required to test a wide range of particles of different physicochemical properties to validate the skin equivalent-human skin particle penetration relationship.

  6. Percutaneous penetration through slightly damaged skin

    DEFF Research Database (Denmark)

    Nielsen, Jesper B

    2005-01-01

    Guidelines for experimental studies of percutaneous penetration prescribe optimal barrier integrity of the skin. The barrier integrity of the skin exposed in occupational or household situations is, however, not always ideal, and skin problems are among the most dominant reasons for absence from ...

  7. Improved Skin Penetration Using In Situ Nanoparticulate Diclofenac Diethylamine in Hydrogel Systems: In Vitro and In Vivo Studies.

    Science.gov (United States)

    Sengupta, Soma; Banerjee, Sarita; Sinha, Biswadip; Mukherjee, Biswajit

    2016-04-01

    Delivering diclofenac diethylamine transdermally by means of a hydrogel is an approach to reduce or avoid systemic toxicity of the drug while providing local action for a prolonged period. In the present investigation, a process was developed to produce nanosize particles (about 10 nm) of diclofenac diethylamine in situ during the development of hydrogel, using simple mixing technique. Hydrogel was developed with polyvinyl alcohol (PVA) (5.8% w/w) and carbopol 71G (1.5% w/w). The formulations were evaluated on the basis of field emission scanning electron microscopy, texture analysis, and the assessment of various physiochemical properties. Viscosity (163-165 cps for hydrogel containing microsize drug particles and 171-173 cps for hydrogel containing nanosize drug particles, respectively) and swelling index (varied between 0.62 and 0.68) data favor the hydrogels for satisfactory topical applications. The measured hardness of the different hydrogels was uniform indicating a uniform spreadability. Data of in vitro skin (cadaver) permeation for 10 h showed that the enhancement ratios of the flux of the formulation containing nanosize drug (without the permeation enhancer) were 9.72 and 1.30 compared to the formulation containing microsized drug and the marketed formulations, respectively. In vivo plasma level of the drug increased predominantly for the hydrogel containing nanosize drug-clusters. The study depicts a simple technique for preparing hydrogel containing nanosize diclofenac diethylamine particles in situ, which can be commercially viable. The study also shows the advantage of the experimental transdermal hydrogel with nanosize drug particles over the hydrogel with microsize drug particles.

  8. Interactions of skin thickness and physicochemical properties of test compounds in percutaneous penetration studies

    NARCIS (Netherlands)

    Wilkinson, S.C.; Maas, W.J.M.; Nielsen, J.B.; Greaves, L.C.; Sandt, J.J.M. van de; Williams, F.M.

    2006-01-01

    Objectives: To determine the effect of skin thickness on the percutaneous penetration and distribution of test compounds with varying physicochemical properties using in vitro systems. Studies were carried out in accordance with OECD guidelines on skin absorption tests. Methods: Percutaneous

  9. Comparison of the Skin Penetration of 3 Metabolically Stable Chemicals Using Fresh and Frozen Human Skin.

    Science.gov (United States)

    Jacques-Jamin, Carine; Duplan, Hélène; Rothe, Helga; Vaillant, Ophelie; Eilstein, Joan; Grégoire, Sebastien; Cubberley, Richard; Lange, Daniela; Ellison, Corie; Klaric, Martina; Hewitt, Nicola; Schepky, Andreas

    2017-01-01

    The Cosmetics Europe ADME Task Force is developing in vitro and in silico tools for predicting skin and systemic concentrations after topical application of cosmetic ingredients. There are conflicting reports as to whether the freezing process affects the penetration of chemicals; therefore, we evaluated whether the storage of human skin used in our studies (8-12 weeks at -20°C) affected the penetration of model chemicals. Finite doses of trans-cinnamic acid (TCA), benzoic acid (BA), and 6-methylcoumarin (6MC) (non-volatile, non-protein reactive and metabolically stable in skin) were applied to fresh and thawed frozen skin from the same donors. The amounts of chemicals in different skin compartments were analysed after 24 h. Although there were some statistical differences in some parameters for 1 or 2 donors, the penetration of TCA, BA, and 6MC was essentially the same in fresh and frozen skin, i.e., there were no biologically relevant differences in penetration values. Statistical differences that were evident indicated that penetration was marginally lower in frozen than in fresh skin, indicating that the barrier function of the skin was not lost. The penetration of the 3 chemicals was essentially unaffected by freezing the skin at -20°C for up to 12 weeks. © 2017 S. Karger AG, Basel.

  10. Is skin penetration a determining factor in skin sensitization ...

    Science.gov (United States)

    Summary:Background. It is widely accepted that substances that cannot penetrate through the skin will not be sensitisers. Thresholds based on relevant physicochemical parameters such as a LogKow > 1 and a MW 1 is a true requirement for sensitisation.Methods. A large dataset of substances that had been evaluated for their skin sensitisation potential, together with measured LogKow values was compiled from the REACH database. The incidence of skin sensitisers relative to non-skin sensitisers below and above the LogKow = 1 threshold was evaluated. Results. 1482 substances with associated skin sensitisation outcomes and measured LogKow values were identified. 305 substances had a measured LogKow penetrate the stratum corneum is a key determinant of skin sensitisation potential and potency. Using the REACH data extracted to test out the validity of common assumptions in the skin sensitization AOP. Builds on trying to develop a proof of concept IATA

  11. Defense against dermal exposures is only skin deep: significantly increased penetration through slightly damaged skin.

    Science.gov (United States)

    Nielsen, Jesper Bo; Nielsen, Flemming; Sørensen, Jens Ahm

    2007-11-01

    The OECD guideline for studies on percutaneous penetration to be used in hazard and risk evaluations prescribes experimental conditions with optimal barrier integrity of the skin, which in many occupational settings probably is not true. Thus, workers may have compromised skin due to chemical or mechanical damage, due to different medical conditions (eczema, dermatitis, skin irritation) or related to occupational scenarios involving prolonged wet work. The present study used the OECD guideline procedures to study the in vitro percutaneous penetration through human skin of a number of model substances (glyphosat, caffeine, benzoic acid, malathion) covering a range of solubilities. Further, we studied the extent to which a slightly damaged skin would change the rate, the amount absorbed during dermal exposure and the distribution of chemical deposition between epidermis and dermis. The present study demonstrates that a limited damage to the skin significantly increases the permeability coefficient (K (p)) as well as total percutaneous penetration of chemicals, and most significantly for those compounds that due to their physicochemical characteristics (the most hydrophilic as well as the most lipophilic) have low penetration rates through intact skin. The present experiment not only confirms the proportionality between lipophilicity and potential for percutaneous penetration, but also illustrates that at a certain degree of lipophilicity of a model compound, the different skin compartments become more attractive for temporary deposition of model compounds. Moreover, a clear change from epidermal deposition towards a dominating dermis deposition of chemicals temporarily deposited within the skin is seen following damage to the skin barrier. Thus, the distribution of chemicals within the skin compartments is affected by the physicochemical characteristics of the chemicals as well as by the integrity of the skin. This observation may have implications when evaluating

  12. An overview of skin penetration enhancers: penetration enhancing activity, skin irritation potential and mechanism of action

    Directory of Open Access Journals (Sweden)

    Sarunyoo Songkro

    2009-08-01

    Full Text Available Transdermal drug delivery has attracted considerable attention over the past 2-3 decades in regard of its many potentialadvantages. However, the role of the skin as a protective barrier renders skin absorption of most drugs problematic. Therefore,skin penetration enhancers are frequently used in the field of transdermal drug delivery in order to reversibly reduce thebarrier function of the stratum corneum, the outermost layer of the skin. To date, a wide range of chemical compounds havebeen shown to enhance the skin penetration of therapeutic drugs. This review presents a critical account of the most commonlyused chemical penetration enhancers (fatty acids and surfactants, and some newer classes of chemical enhancers (terpenes,polymers, monoolein, oxazolidinones, with emphasis on their efficacy, mechanism of action, and skin irritation potential. Thisreview also discusses the traditional and more recently developed methods for the screening and evaluation of chemical penetration enhancers, and addresses the continuing problems in the rational selection of a chemical penetration enhancer for a specific drug to be delivered via the transdermal route.

  13. Noninvasive penetration of 5 nm hyaluronic acid molecules across the epidermal barrier (in vitro) and its interaction with human skin cells.

    Science.gov (United States)

    Nashchekina, Yu A; Raydan, M

    2018-02-01

    Hyaluronic acid represents one of the major components of the extracellular environment. The main challenge remains in the ability to deliver these molecules noninvasively across the skin barrier, which can be overcome by the reduction in size to an extent that allows these molecules to pass across the skin barrier. The aim of this study was to measure the penetration and bioavailability of low molecular weight hyaluronic acid to cross an epidermal barrier model. Determining the quantity of hyaluronic acid in the test solutions was carried with method of photocolorimetry analysis. Investigation of the interaction of cells with LMWHA was studied with a confocal microscope. The study showed that LMWHA is able to cross the epidermis. Most effective penetration level is during the first 6 hours reaching 75%, and then the concentration started to decline and reached the equilibrium state within the following 2 hours. Confocal laser microscopy demonstrated different distribution and behavior of these molecules among the keratinocytes and fibroblasts. Reducing the size of hyaluronic acid to 5 nm enhance their transport across the epidermal layer. The concentration of hyaluronic acid molecules was higher on the fibroblast surface in comparison to their extracellular environment. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Skin Penetration Enhancement by Natural Oils for Dihydroquercetin Delivery.

    Science.gov (United States)

    Čižinauskas, Vytis; Elie, Nicolas; Brunelle, Alain; Briedis, Vitalis

    2017-09-12

    Natural oils are commonly used in topical pharmaceutical formulations as emulsifiers, stabilizers or solubility enhancers. They are presented as safe and inert components, mainly used for formulation purposes. It is confirmed that natural oils can affect the skin penetration of various substances. Fatty acids are mainly responsible for this effect. Current understanding lacks reliable scientific data on penetration of natural oils into the skin and their skin penetration enhancement potential. In the current study, fatty acid content analysis was used to determine the principal fatty acids in soybean, olive, avocado, sea-buckthorn pulp, raspberry seed and coconut oils. Time of flight secondary ion mass spectrometry bioimaging was used to determine the distribution of these fatty acids in human skin ex vivo after application of the oils. Skin penetration enhancement ratios were determined for a perspective antioxidant compound dihydroquercetin. The results demonstrated skin penetration of fatty acids from all oils tested. Only soybean and olive oils significantly increased the skin distribution of dihydroquercetin and can be used as skin penetration enhancers. However, no correlation can be determined between the fatty acids' composition and skin penetration enhancement using currently available methodological approaches. This indicates that potential chemical penetration enhancement should be evaluated during formulation of topically applied products containing natural oils.

  15. Dermal In Vitro Penetration of Methiocarb, Paclobutrazol, and Pirimicarb: Effect of Nonylphenolethoxylate and Protective Gloves

    National Research Council Canada - National Science Library

    Jesper Bo Nielsen; Helle Raun Andersen

    2001-01-01

    .... In an experiment using in vitro static diffusion cells mounted with human skin, we evaluated the effect of nonylphenolethoxylate on dermal penetration of three extensively used pesticides-methiocarb...

  16. Age-related percutaneous penetration part 1: skin factors.

    Science.gov (United States)

    Konda, S; Meier-Davis, S R; Cayme, B; Shudo, J; Maibach, H I

    2012-05-01

    Changes in the skin that occur in the elderly may put them at increased risk for altered percutaneous penetration from pharmacotherapy along with potential adverse effects. Skin factors that may have a role in age-related percutaneous penetration include blood flow, pH, skin thickness, hair and pore density, and the content and structure of proteins, glycosaminoglycans (GAGs), water, and lipids. Each factor is examined as a function of increasing age along with its potential impact on percutaneous penetration. Additionally, topical drugs that successfully overcome the barrier function of the skin can still fall victim to cutaneous metabolism, thereby producing metabolites that may have increased or decreased activity. This overview discusses the current data and highlights the importance of further studies to evaluate the impact of skin factors in age-related percutaneous penetration.

  17. Liposome surface charge influence on skin penetration behaviour.

    Science.gov (United States)

    Gillet, A; Compère, P; Lecomte, F; Hubert, P; Ducat, E; Evrard, B; Piel, G

    2011-06-15

    Vesicular systems have shown their ability to increase dermal and transdermal drug delivery. Their mechanism of drug transport into and through the skin has been investigated but remains a much debated question. Several researchers have outlined that drug penetration can be influenced by modifying the surface charge of liposomes. In the present work we study the influence of particle surface charge on skin penetration. The final purpose is the development of a carrier system which is able to enhance the skin delivery of two model drugs, betamethasone and betamethasone dipropionate. Liposomes were characterised by their size, morphology, zeta potential, encapsulation efficiency and stability. Ex vivo diffusion studies using Franz diffusion cells were performed. Confocal microscopy was performed to visualise the penetration of fluorescently labelled liposomes into the skin. This study showed the potential of negatively charged liposomes to enhance the skin penetration of betamethasone and betamethasone dipropionate. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Effect of sodium lauryl sulfate (SLS) on in vitro percutaneous penetration of water, hydrocortisone and nickel

    DEFF Research Database (Denmark)

    Frankild, S; Andersen, Klaus Ejner; Nielsen, Gunnar

    1995-01-01

    The dose- and time-related effect of sodium lauryl sulfate (SLS) on in vitro percutaneous penetration was studied using 3 radiolabeled tracer compounds with different physicochemical properties: tritiated water, hydrocortisone and nickel. Human cadaver abdominal skin from caucasian women was used....../damaging effect on the skin barrier. It should be kept in mind that the model uses a dead skin membrane without the barrier repair mechanisms of live skin....

  19. Interactions of skin thickness and physicochemical properties of test compounds in percutaneous penetration studies

    DEFF Research Database (Denmark)

    Wilkinson, Simon C.; Maas, Wilfred J. M.; Nielsen, Jesper Bo

    2006-01-01

    Abstract   Objectives: To determine the effect of skin thickness on the percutaneous penetration and distribution of test compounds with varying physicochemical properties using in vitro systems. Studies were carried out in accordance with OECD guidelines on skin absorption tests. Methods......: Percutaneous penetration of caffeine (log P -0.01), testosterone (log P 3.32), propoxur (log P 1.52) (finite dose in ethanol to water vehicle ratio) and butoxyethanol (log P 0.83) (undiluted finite dose or as an infinite dose 50% [v/v] aqueous solution) through skin of varying thicknesses under occluded......: The maximum flux of caffeine was increased with decreasing skin thickness, although these differences were found to be non-significant. The presence of caffeine in the skin membrane was not altered by skin thickness. Maximum flux and cumulative dose absorbed of testosterone and butoxyethanol (in both finite...

  20. Release of rosmarinic acid from semisolid formulations and its penetration through human skin ex vivo

    Directory of Open Access Journals (Sweden)

    Stelmakienė Ada

    2015-06-01

    Full Text Available The aim of this study was to evaluate the release of rosmarinic acid (RA from the experimental topical formulations with the Melissa officinalis L. extract and to evaluate its penetration through undamaged human skin ex vivo. The results of the in vitro release study showed that higher amounts of RA were released from the emulsion vehicle when lemon balm extract was added in its dry form. An inverse correlation was detected between the released amount of RA and the consistency index of the formulation. Different penetration of RA into the skin may be influenced by the characteristics of the vehicle as well as by the form of the extract. The results of penetration assessment showed that the intensity of RA penetration was influenced by its lipophilic properties: RA was accumulating in the epidermis, while the dermis served as a barrier, impeding its deeper penetration.

  1. In vitro evaluation of copaiba oil as a kojic acid skin enhancer

    National Research Council Canada - National Science Library

    Robson Vicente Machado de Oliveira; Mitsuko Taba Ohara; Marta Maria Duarte Carvalho Vila; Marcos Moisés Gonçalves

    2010-01-01

    The capacity of copaíba oil to act as a skin penetration enhancer for the depigmenting agent kojic acid was evaluated using an in vitro diffusion system with static flux and shed rattlesnake skin membrane, Crotalus...

  2. Evaluation of nicotinamide microemulsion on the skin penetration enhancement.

    Science.gov (United States)

    Boonme, Prapaporn; Boonthongchuay, Chalida; Wongpoowarak, Wibul; Amnuaikit, Thanaporn

    2016-01-01

    This study purposed to evaluate a microemulsion containing nicotinamide for its characteristics, stability, and skin penetration and retention comparing with a solution of nicotinamide in 2:1 mixture of water and isopropyl alcohol (IPA). The microemulsion system was composed of 1:1 mixture of Span80 and Tween80 as a surfactant mixture, isopropyl palmitate (IPP) as an oil phase, and 2:1 mixture of water and IPA as an aqueous phase. Nicotinamide microemulsion was prepared by dissolving the active in the aqueous phase before simply mixing with the other components. It was determined for its characteristics and stability under various conditions. The skin penetration and retention studies of nicotinamide microemulsion and solution were performed by modified Franz diffusion cells, using newborn pig skin as the membrane. The results showed that nicotinamide microemulsion could be obtained as clear yellowish liquid, was water-in-oil (w/o) type, possessed Newtonian flow, and exhibited physicochemical stability when kept at 4 °C and room temperature (≈30 ± 2 °C) during 3 months. From the skin penetration data, the microemulsion could enhance the skin penetration of nicotinamide comparing with the solution. Additionally, nicotinamide microemulsion could provide much higher amount of skin retention than that of skin penetration, resulting in suitability for a cosmeceutical product.

  3. Dermal in vitro penetration of methiocarb, paclobutrazol, and pirimicarb.

    Science.gov (United States)

    Nielsen, J B; Nielsen, F

    2000-11-01

    The dominant route of occupational exposure to pesticides in horticulture is dermal. However, preventive measures are seldom used when handling plant cultures recently treated with pesticides, thus causing significant dermal exposure and potential absorption. Assessment of exposure often depends on biological monitoring of blood or urine samples. The skin often acts as a temporary reservoir for chemicals before absorption. Failure to consider the lag time between dermal exposure and appearance of pesticide or metabolites in the general circulation may lead to false conclusions about assessment of exposure. In an experimental model in which in vitro static diffusion cells were mounted with human skin, dermal penetration of three extensively used pesticides (methiocarb, paclobutrazol, pirimicarb) was evaluated. Pirimicarb and paclobutrazol had comparable rates of dermal penetration and lag times of around 18 hours. Methiocarb had a considerably shorter lag time. Dermal penetration continued for extended periods after exposure had ended. With lag times sometimes considerably longer than a normal working day, biological monitoring at the end of exposure may seriously underestimate the actual exposure. There may be implications for regulatory guidelines, which often require only 24 hour observation periods.

  4. Measuring the penetration of a skin sensitizer and its delivery vehicles simultaneously with confocal Raman spectroscopy.

    Science.gov (United States)

    Bonnist, E Y M; Gorce, J-P; Mackay, C; Pendlington, R U; Pudney, P D A

    2011-01-01

    Among the factors determining the propensity of a chemical to induce skin allergy are the penetration into skin and the kinetics of ingress. Confocal Raman spectroscopy can provide such information as it enables direct, spatially resolved measurement of the skin and of any chemical uptake. Several chemicals can be monitored at once, and the method is non-destructive (light in, light out) so that the skin can be kept intact for repeated and continuous measurement. Raman spectroscopy was used to follow the penetration of 2.5 weight percent trans-cinnamaldehyde and its delivery vehicle into skin in vitro, up to 24 h after topical application. A custom-made Bronaugh-type diffusion cell that was suitable for the Raman experiment was used. Four different vehicles were tested: absolute ethanol, 50% aqueous ethanol, propylene glycol and acetone:olive oil (4:1); these gave different time scales for cinnamaldehyde penetration. The acetone:olive oil vehicle phase-separated on the skin surface and the cinnamaldehyde penetrated at different rates in the different phases, which may be of significance since this is the preferred solvent for the local lymph node assay (an in vivo animal test used to generate hazard information on skin sensitization). In conclusion, the Raman method gives valuable detailed information on chemical ingress, clearly differentiates between different delivery rates and allows solvent monitoring alongside the chemical of interest. Copyright © 2011 S. Karger AG, Basel.

  5. Development of a stratum corneum substitute for in vitro percutaneous penetration studies : a skin barrier model comprising synthetic stratum corneum lipids

    NARCIS (Netherlands)

    Jager, Miranda Wilhelmina de

    2006-01-01

    The research outlined in this thesis was focused on the development of a skin barrier model, which can substitute for stratum corneum in diffusion studies. This so-called stratum corneum substitute (SCS) was prepared with reconstituted SC lipids (cholesterol, free fatty acids and ceramides) on a

  6. Dermal in vitro penetration of methiocarb, paclobutrazol, and pirimicarb: effect of nonylphenolethoxylate and protective gloves.

    Science.gov (United States)

    Nielsen, J B; Andersen, H R

    2001-02-01

    Dermal exposure has become the major route of human occupational exposure to pesticides. Detergents are used as part of formulated pesticide products and are known to change the barrier properties of human skin in vitro. However, studies on the influence of detergents as well as protective glove materials on dermal penetration of pesticides are scarce. In an experiment using in vitro static diffusion cells mounted with human skin, we evaluated the effect of nonylphenol-ethoxylate on dermal penetration of three extensively used pesticides--methiocarb, paclobutrazol, and pirimicarb--and the protection against dermal penetration offered by protective gloves made of latex or nitrile. There was a general tendency, though not statistically significant for all pesticides, for nonylphenolethoxylate to decrease the percutaneous penetration of the three pesticides. The nitrile generally offered better protection against percutaneous penetration of pesticides than did latex, but the degree of protection decreased over time and depended on the pesticides used.

  7. Adapalene pretreatment increases follicular penetration of clindamycin: In vitro and in vivo studies

    Directory of Open Access Journals (Sweden)

    Jain Gaurav

    2007-01-01

    Full Text Available Background: Topical retinoids normalize desquamation, reduce comedogenesis and may enhance the penetration of other topicals providing more effective treatment of acne. Aim: We evaluated the effect of adapalene on skin penetration of clindamycin phosphate when it is applied concomitantly or after various time durations following adapalene application. Methods: The in vitro studies were carried out using excised rat skin, whereas the in vivo studies were conducted on healthy human volunteers. Radioactive clindamycin phosphate (1% gel was applied to rat skin sections and to the hands of human volunteers concomitantly and after the pretreatment of the skin for 3, 5 and 10 min with 10 mg of adapalene (0.1% gel. Quantification of clindamycin phosphate was performed by liquid scintillation. Results: In vitro skin penetration and distribution of clindamycin phosphate was affected by the pretreatment time. Significantly higher skin concentration of clindamycin phosphate (15.5% with largest proportion in viable skin layer (9.4% of applied dose was found when clindamycin phosphate gel was applied after the pretreatment of the skin with adapalene gel for 5 min. Further increase in pretreatment time has no additive influence on the penetration of clindamycin phosphate. In vivo results were in corroboration with the in vitro results and demonstrate significantly higher concentration of clindamycin phosphate (19% in the skin following pretreatment with adapalene gel for 5 min. Adapalene acts as a penetration enhancer and increases the penetration of topical clindamycin phosphate. Conclusion: Application of clindamycin phosphate gel after the pretreatment of skin with adapalene gel for 5 min may contribute significantly to the increased efficacy of therapy.

  8. Skin penetration of silicon dioxide microneedle arrays.

    Science.gov (United States)

    Kim, Sangchae; Shetty, S; Price, D; Bhansali, S

    2006-01-01

    Out-of-plane hollow silicon dioxide microneedle arrays were fabricated and investigated to determine their efficacy for transdermal applications. The fabrication process of the SiO2 microneedles is described, and mechanical fracture forces were investigated on microneedles with different geometrical dimensions. Biomechanical characterization of the microneedles was performed to specifically test for reliable stratum corneum and skin insertion by changing the regulatory parameters such as needle width and cross-section.

  9. Pharmacokinetics and skin-tissue penetration of daptomycin in rats

    Directory of Open Access Journals (Sweden)

    Matsumoto K

    2015-05-01

    Full Text Available Kazuaki Matsumoto,1 Masashi Kitaoka,1 Yuko Kuroda,1 Kazuro Ikawa,2 Norifumi Morikawa,2 Junichi Sasaki,3 Osamu Iketani,4 Satoshi Iwata,4 Tetsuya Horino,5 Seiji Hori,5 Junko Kizu1 1Division of Practical Pharmacy, Keio University Faculty of Pharmacy, Tokyo, 2Department of Clinical Pharmacotherapy, Hiroshima University, Hiroshima, 3Department of Emergency and Critical Care Medicine, 4Center for Infectious Diseases and Infection Control, Keio University School of Medicine, 5Department of Infectious Diseases and Infection Control, Jikei University School of Medicine, Tokyo, Japan Background: Daptomycin is recommended for complicated skin and skin-structure infections. However, information on the penetration of daptomycin into skin is limited. Therefore, the aim of this in vivo investigation was to determine the pharmacokinetics and skin penetration of daptomycin in rats. Materials and methods: Concentrations of daptomycin were determined by high-performance liquid chromatography. A noncompartmental pharmacokinetic analysis was conducted to estimate the rate and extent of daptomycin penetration from the systemic circulation into skin tissue. Since protein binding of daptomycin in rat serum was 89.3%, the free maximum concentration (Cmax and free area under the curve from time 0 to infinity (AUC0–∞ for plasma were calculated as follows: fCmax, plasma = (1 – 0.893 × Cmax, plasma, fAUC0–∞, plasma = (1 – 0.893 × AUC0–∞, plasma. Results: The following values (mean ± standard deviation were obtained: 0.06±0 L/h/kg for total clearance (CLtotal, 0.44±0.06 hours for elimination-rate constant, 1.58±0.23 hours for half-life, 0.14±0.02 L/kg for steady-state volume distribution, and 2.28±0.33 hours for mean residence time. Time to Cmax was 3.0 hours for plasma and skin tissue. Cmax and AUC0–∞ for plasma were 175.8±5.1 µg/mL and 811.8±31.9 µg × h/mL, respectively. Cmax and AUC0–∞ for skin tissue were 19.1±1.7 µg/mL and 113.9

  10. Effect of Different Skin Penetration Promoters in Halobetasol Propionate Permeation and Retention in Human Skin

    Directory of Open Access Journals (Sweden)

    Paulina Carvajal-Vidal

    2017-11-01

    Full Text Available Halobetasol propionate (HB is a potent synthetic corticosteroid used against inflammatory skin diseases, such as dermatitis, eczema, and psoriasis, among others. The aim of this study is to define how the presence of different skin penetration enhancers (nonane, menthone, limonene, azone, carene, decanol, linoleic acid and cetiol affects the penetration and retention in skin of HB. To determine drug penetration through skin, 5% of each promoter was used in an ex vivo system with human skin on Franz cells. The results showed that the highest permeation occurs in the presence of menthone, followed by nonane. Permeation parameters were determined. The in vivo test was assessed, and the formulation containing HB-menthone presented better anti-inflammatory efficacy. These results are useful to generate a specific treatment according to each patient’s needs, and the inflammatory characteristics of the disease.

  11. Development of a HPLC method for determination of four UV filters in sunscreen and its application to skin penetration studies.

    Science.gov (United States)

    Souza, Carla; Maia Campos, Patrícia M B G

    2017-12-01

    This study describes the development, validation and application of a high-performance liquid chromatography (HPLC) method for the simultaneous determination of the in vitro skin penetration profile of four UV filters on porcine skin. Experiments were carried out on a gel-cream formulation containing the following UV filters: diethylamino hydroxybenzoyl hexyl benzoate (DHHB), bis-ethylhexyloxyphenol methoxyphenyl triazine (BEMT), methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT) and ethylhexyl triazone (EHT). The HPLC method demonstrated suitable selectivity, linearity (10.0-50.0 μg/mL), precision, accuracy and recovery from porcine skin and sunscreen formulation. The in vitro skin penetration profile was evaluated using Franz vertical diffusion cells for 24 h after application on porcine ear skin. None of the UV filters penetrated the porcine skin. Most of them stayed on the skin surface (>90%) and only BEMT, EHT and DHHB reached the dermis plus epidermis layer. These results are in agreement with previous results in the literature. Therefore, the analytical method was useful to evaluate the in vitro skin penetration of the UV filters and may help the development of safer and effective sunscreen products. Copyright © 2017 John Wiley & Sons, Ltd.

  12. Tretinoin-based formulations - influence of concentration and vehicles on skin penetration

    Directory of Open Access Journals (Sweden)

    Edileia Bagatin

    2015-03-01

    Full Text Available Tretinoin is used in the management of acne and it is part of a gold standard treatment for photoaging. It has also been reported as an agent for superficial chemical peeling in highly concentrated formulations with few considerations about skin penetration. The aim of this study was to evaluate the influence of drug concentration and vehicles currently used on skin penetration of tretinoin. In vitro permeation tests were carried out using Franz diffusion cells fitted with porcine ear skin and 10% aqueous methanol in the receptor compartment. Formulations studied, cream or hydroalcoholic dispersion, containing 0.25%, 1% and 5% of tretinoin were placed in the donor compartment for six hours. Tretinoin concentration in skin layers was measured by high performance liquid chromatography. The largest amount of tretinoin from both vehicles was detected in stratum corneum with significant differences among the three concentrations. The hydroalcoholic dispersion was the best vehicle. Significant amounts of tretinoin were found even in deep layers of epidermis. The formulation with 0.25% tretinoin showed better results when considered the amount of tretinoin on skin in terms of percentage. Finally, skin penetration of tretinoin was influenced by vehicle and concentration of this drug used in formulation.

  13. In vitro percutaneous penetration and characterization of silver from silver-containing textiles

    NARCIS (Netherlands)

    Bianco, Carlotta; Kezic, Sanja; Crosera, Matteo; Svetličić, Vesna; Šegota, Suzana; Maina, Giovanni; Romano, Canzio; Larese, Francesca; Adami, Gianpiero

    2015-01-01

    The objective of this study was to determine the in vitro percutaneous penetration of silver and characterize the silver species released from textiles in different layers of full thickness human skin. For this purpose, two different wound dressings and a garment soaked in artificial sweat were

  14. A custom tailored model to investigate skin penetration in porcine skin and its comparison with human skin.

    Science.gov (United States)

    Herbig, Michael E; Houdek, Pia; Gorissen, Sascha; Zorn-Kruppa, Michaela; Wladykowski, Ewa; Volksdorf, Thomas; Grzybowski, Stephan; Kolios, Georgios; Willers, Christoph; Mallwitz, Henning; Moll, Ingrid; Brandner, Johanna M

    2015-09-01

    Reliable models for the determination of skin penetration and permeation are important for the development of new drugs and formulations. The intention of our study was to develop a skin penetration model which (1) is viable and well supplied with nutrients during the period of the experiment (2) is mimicking human skin as far as possible, but still is independent from the problems of supply and heterogeneity, (3) can give information about the penetration into different compartments of the skin and (4) considers specific inter-individual differences in skin thickness. In addition, it should be quick and inexpensive (5) and without ethical implications (6). Using a chemically divers set of four topically approved active pharmaceutical ingredients (APIs), namely diclofenac, metronidazole, tazarotene, and terbinafine, we demonstrated that the model allows reliable determination of drug concentrations in different layers of the viable epidermis and dermis. For APIs susceptible for skin metabolism, the extent of metabolic transformation in epidermis and dermis can be monitored. Furthermore, a high degree of accordance in the ability for discrimination of skin concentrations of the substances in different layers was found in models derived from porcine and human skin. Viability, proliferation, differentiation and markers for skin barrier function were surveyed in the model. This model, which we call 'Hamburg model of skin penetration' is particularly suited to support a rational ranking and selection of dermatological formulations within drug development projects. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Sharpening of hollow silicon microneedles to reduce skin penetration force

    Science.gov (United States)

    Khanna, Puneet; Luongo, Kevin; Strom, Joel A.; Bhansali, Shekhar

    2010-04-01

    In this research, hollow silicon microneedles with sharpened tips have been fabricated without any reduction to the needle shaft diameter. By sharpening the needles only at the tip and not over the entire length of the needle, their mechanical strength is maintained, while reducing the insertion force into skin. The process achieves this geometry by novel use photoresist depletion during DRIE. Microneedles of varying levels of tip sharpness were tested on human cadaver skin to measure their force of penetration. The results show a marked decrease of insertion force with progressive sharpening of microneedle tips, reducing more than 75 times in magnitude for extremely sharp tips. The toughness of human skin was derived to be approximately 24.28 kJ m-2.

  16. Interaction of sunscreen TiO2 nanoparticles with skin and UV light: penetration, protection, phototoxicity

    Science.gov (United States)

    Popov, Alexey; Lademann, Jürgen; Priezzhev, Alexander; Myllylä, Risto

    2009-07-01

    Titanium dioxide (TiO2) nanoparticles are extensively used nowadays in sunscreens as protective compounds for human skin from UV radiation. In this paper, such particles are investigated from the viewpoint of penetration into living skin, UV protective properties (compared with silicon (Si) particles) and as sources of free radicals if UV-irradiated. We show that: a) even after multiple applications, the particles are located within the uppermost 3-μm-thick part of the skin; b) the optimal sizes are found to be 62 nm and 55 nm, respectively for TiO2 and Si particles for 310-nm light and, correspondingly, 122 and 70 nm - for 400-nm radiation; c) if applied onto glass, small particles of 25 nm in diameter produce an increased amount of free radicals compared to the larger ones of 400 nm in diameter and placebo itself; however, if applied onto porcine skin in vitro, there is no statistically distinct difference in the amount of radicals generated by the two kinds of particles on skin and by the skin itself. This proves that although particles as part of sunscreens produce free radicals, the effect is negligible in comparison to the production of radicals by skin in vitro.

  17. Penetration and growth of DPPC/DHPC bicelles inside the stratum corneum of the skin.

    Science.gov (United States)

    Barbosa-Barros, L; de la Maza, A; Estelrich, J; Linares, A M; Feliz, M; Walther, P; Pons, R; López, O

    2008-06-03

    The effect of dipalmitoyl phosphatidylcholine (DPPC)/dihexanoyl phosphatidylcholine (DHPC) bicelles on the microstructure of pig stratum corneum (SC) in vitro was evaluated. The physicochemical characterization of these nanoaggregates revealed small disks with diameters around 15 nm and a thickness of 5.4 nm. Upon dilution, the bicelles grow and transform into vesicles. Cryogenic scanning electron microscopy (cryo-SEM) images of the SC pieces treated with this system showed vesicles of about 200 nm and lamellar-like structures in the intercellular lipid areas. These vesicles probably resulted from the growth and molecular rearrangement of the DPPC/DHPC bicelles after penetrating the SC. The presence of lamellar-like structures is ascribed to the interaction of the lipids from bicelles with the SC lipids. The bicellar system used is suitable to penetrate the skin SC and to reinforce the intercellular lipid areas, constituting a promising tool for skin applications.

  18. Measuring skin penetration by confocal Raman microscopy (CRM): correlation to results from conventional experiments

    Science.gov (United States)

    Lunter, Dominique; Daniels, Rolf

    2016-03-01

    Confocal Raman microscopy has become an advancing technique in the characterization of drug transport into the skin. In this study the skin penetration of a local anesthetic from a semisolid preparation was investigated. Furthermore, the effect of the chemical enhancers propylene glycol and POE-23-lauryl ether on its penetration was investigated. The results show that confocal Raman microscopy may provide detailed information on the penetration of APIs into the skin and may elucidate their distribution within the skin with high resolution. The results of the CRM analysis are fully in line with those of conventional permeation and penetration experiments.

  19. Alkylglycerol Derivatives, a New Class of Skin Penetration Modulators

    Directory of Open Access Journals (Sweden)

    Sergio Alberto Bernal-Chávez

    2017-01-01

    Full Text Available The absorption modulating activity of two alkylglycerol derivatives (batyl and chimyl alcohol on skin barrier properties was evaluated. Biophysical tests such as transepidermal water loss (TEWL and attenuated total reflectance–Fourier transform infrared (ATR-FTIR spectroscopy, as well as in vitro skin permeation studies, were performed in order to determine the effect of these compounds as chemical absorption modulators. Four drugs were used as models: three NSAIDS (diclofenac, naproxen, and piroxicam and glycyrrhizic acid. The results showed that treatment of the skin with alkylglycerols caused (i a reduction on the amount of drug permeated; (ii a reduction in TEWL; and (iii changes in the ATR-FTIR peaks of stratum corneum lipids, indicative of a more ordered structure. All of these findings confirm that alkyl glycerols have an absorption retarding effect on the drugs tested. Such effects are expected to give rise to important applications in the pharmaceutical and cosmetic sectors, in cases where it is desirable for the drug to remain in the superficial layers of the skin to achieve a local effect.

  20. Quantification of quantum dot murine skin penetration with UVR barrier impairment

    Science.gov (United States)

    Mortensen, Luke J.; Jatana, Samreen; Gelein, Robert; De Benedetto, Anna; De Mesy Bentley, Karen L.; Beck, Lisa; Elder, Alison; DeLouise, Lisa A.

    2013-01-01

    Ultraviolet radiation (UVR) skin exposure is a common exogenous insult that can alter skin barrier and immune functions. With the growing presence of nanoparticles (NPs) in consumer goods and technological applications the potential for NPs to contact UVR exposed skin is increasing. Therefore it is important to understand the effect of UVR on NP skin penetration and potential for systemic translocation. Previous studies qualitatively showed that UVR skin exposure can increase the penetration of NPs below the stratum corneum. In the present work, an in vivo mouse model was used to quantitatively examine the skin penetration of carboxylated (CdSe/ZnS, core/shell) quantum dots (QDs) through intact and UVR barrier disrupted murine skin by organ Cd mass analysis. Transepidermal water loss was used to measure the magnitude of the skin barrier defect as a function of dose and time post UVR exposure. QDs were applied to mice 3-4 days post UVR exposure at the peak of the skin barrier disruption. Our results reveal unexpected trends that suggest these negative charged QDs can penetrate barrier intact skin and that penetration and systemic transport depends on the QD application time post UVR exposure. The effect of UVR on skin resident dendritic cells and their role in the systemic translocation of these QDs are described. Our results suggest that NP skin penetration and translocation may depend on the specific barrier insult and the inflammatory status of the skin. PMID:23078247

  1. Natural oils affect the human skin integrity and the percutaneous penetration of benzoic acid dose-dependently

    DEFF Research Database (Denmark)

    Nielsen, Jesper Bo

    2006-01-01

    Abstract: Natural oils are extensively used in cosmetics and as treatment for a growing number of more or less specific ailments. Skin irritation and cases of allergy have repeatedly been described in the literature following exposure to these oils. The present study evaluated the extent to which...... three natural oils (eucalyptus oil, tea tree oil, peppermint oil) would affect the skin integrity and the percutaneous penetration of benzoic acid when applied topically in relevant concentrations. An experimental in vitro model using static diffusion cells mounted with human breast or abdominal skin...... was applied. The three natural oils decreased the skin integrity dose-dependently. Concomitant dermal exposure to low concentrations of peppermint oil reduced the percutaneous penetration of benzoic acid. The present study lends support to the notion that low concentrations of peppermint oil may act...

  2. In vitro and ex vivo corneal penetration and absorption models.

    Science.gov (United States)

    Agarwal, Priyanka; Rupenthal, Ilva D

    2016-12-01

    Topical drug administration is the preferred route of drug delivery to the eye despite the poor bioavailability. To develop more efficient drug carriers, reliable in vitro or ex vivo models are required in the early stages of formulation development, with such methods being faster, cheaper, and more ethical alternatives to in vivo studies. In vitro cell culture models are increasingly being used for transcorneal penetration studies, with primary cell cultures and immortalized cell lines now giving way to the development of organotypic corneal constructs for studying ocular drug bioavailability. Artificially cultured human corneal equivalents are still in the early stages of development, but present tremendous potential for corneal penetration studies. Ex vivo models using excised animal tissue are also being used to study corneal penetration with promising results, although significant inter-species variations need to be considered. This review discusses the in vitro and ex vivo models currently being used to study corneal penetration and evaluates their advantages and limitations with a focus on diffusion cell assemblies. In addition to the tissue used, the diffusion cell set-up can significantly influence the penetration profile and should be cautiously adjusted to simulate clinical conditions.

  3. Impact of semi-solid formulations on skin penetration of iron oxide nanoparticles.

    Science.gov (United States)

    Musazzi, Umberto M; Santini, Benedetta; Selmin, Francesca; Marini, Valentina; Corsi, Fabio; Allevi, Raffaele; Ferretti, Anna M; Prosperi, Davide; Cilurzo, Francesco; Colombo, Miriam; Minghetti, Paola

    2017-02-17

    This work aimed to provide useful information on the incidence of the choice of formulation in semi-solid preparations of iron-oxide nanoparticles (IONs). The appropriate analytical methods to assess the IONs physical stability and the effect of the semi-solid preparations on IONs human skin penetration were discussed. The physical stability of IONs (Dh = 31 ± 4 nm; ζ = -65 ± 5 mV) loaded in five semi-solid preparations (0.3% w/v), namely Carbopol gel (CP), hydroxyethyl cellulose gel (HEC), carboxymethylcellulose gel (CMC), cetomacrogol cream (Cet) and cold cream was assessed by combining DLS and low-field pulsed NMR data. The in vitro penetration of IONs was studied using human epidermis or isolated stratum corneum (SC). Reversible and irreversible IONs aggregates were evidenced only in HEC and CMC, respectively. IONs diffused massively through SC preferentially by an intercellular pathway, as assessed by transmission electron microscopy. The semi-solid preparations differently influenced the IONs penetration as compared to the aqueous suspension. Cet cream allowed the highest permeation and the lowest retained amount, while cold cream and CP favored the accumulation into the skin membrane. Basic cutaneous semi-solid preparations could be used to administer IONs without affecting their permeation profile if they maintained their physical stability over time. This property is better discriminated by low-field pulsed NMR measurements than the commonly used DLS measurements.

  4. Hydrogen sulphide and phosphine interactions with human skin in vitro.

    Science.gov (United States)

    Gaskin, Sharyn; Heath, Linda; Pisaniello, Dino; Evans, Richard; Edwards, John W; Logan, Michael; Baxter, Christina

    2017-04-01

    Accidental or intentional releases of toxic gases can have significant public health consequences and emergency resource demands. Management of exposed individuals during hazardous material incidents should be risk and evidence based, but there are knowledge gaps in relation to dermal absorption of gases and management advice for potentially exposed individuals. Using a modified Organization for Economic Co-operation and Development (OECD) in vitro toxicology protocol with human donor skin, this article reports on two common and odorous chemicals, hydrogen sulphide and phosphine. Results show that undamaged human skin provides a good barrier to hydrogen sulphide (up to 800 ppm) and phosphine (up to 1000 ppm) penetration for up to 30 min exposures, with little variability in the presence of clothing or in elevated temperature and humidity conditions. A practical guideline template for skin decontamination has been developed, and implications of the research for first responders are outlined.

  5. Cutaneous Permeation and Penetration of Sunscreens: Formulation Strategies and In Vitro Methods

    Directory of Open Access Journals (Sweden)

    Silvia Tampucci

    2017-12-01

    Full Text Available Sunscreens are the most common products used for skin protection against the harmful effects of ultraviolet radiation. However, as frequent application is recommended, the use of large amount of sunscreens could reflect in possible systemic absorption and since these preparations are often applied on large skin areas, even low penetration rates can cause a significant amount of sunscreen to enter the body. An ideal sunscreen should have a high substantivity and should neither penetrate the viable epidermis, the dermis and the systemic circulation, nor in hair follicle. The research of methods to assess the degree of penetration of solar filters into the skin is nowadays even more important than in the past, due to the widespread use of nanomaterials and the new discoveries in cosmetic formulation technology. In the present paper, different in vitro studies, published in the last five years, have been reviewed, in order to focus the attention on the different methodological approaches employed to effectively assess the skin permeation and retention of sunscreens.

  6. Aircraft Skin Penetrator and Agent Applicator. Volume 2. Test and Evaluation.

    Science.gov (United States)

    1984-11-01

    which will penetrate aircraft skin and serve as a discharge outlet to dispense fire - extinguishing agent. AMETEK, Inc./ORED designed a tool to meet...grease Degrease with 1,1,1-Trichloro- ethane, steem clean and blow dry. 3. Odor Rinse with solution of baking soda (sodium bicarbonate) then rinse with...development and construction of an aircraft skin penetrator device to provide rapid penetration and allow placement of a suitable fire suppressing agent onto

  7. In vitro evaluation of cutaneous penetration of acyclovir from semisolid commercial formulations and relation with its effective antiviral concentration

    Directory of Open Access Journals (Sweden)

    Rafaela Martins Sponchiado

    Full Text Available ABSTRACT The evaluation of drug permeation/penetration of semisolid formulations into animal skin can be useful to supplement the pharmaceutical equivalence. This paper describes the in vitro assessment of acyclovir (ACV into porcine skin from commercial formulations with etermination of drug concentration in different layers of cutaneous tissue to correlate with effective antiviral concentration in order to improve the equivalence decision. Studies were conducted using Franz cells and porcine skin. Selected pharmaceutical creams containing ACV had identical (reference and generic and different (similar excipients. A software program was employed for the simulation of antiviral effectiveness in the skin. Regarding ACV skin penetration, the first batch of the generic product showed a significant difference from reference and similar products, while in the second batch all products demonstrated equivalent drug penetration in the skin. Simulation studies suggest that formulations analysed exhibit a pharmacological effect even when in contact with Herpes simplex strains of high IC50 (inhibitory concentration required to reduce viral replication by 50%. According to results, it can be assumed that the in vitro cutaneous permeation/penetration study does not supply sensitivity information regarding small alterations of ACV semisolid formulations due to the variability inherent to the method, although it can be relevant to pharmaceutical equivalence studies in the development of semisolid products.

  8. Measurement of skin permeation/penetration of nanoparticles for their safety evaluation.

    Science.gov (United States)

    Kimura, Eriko; Kawano, Yuichiro; Todo, Hiroaki; Ikarashi, Yoshiaki; Sugibayashi, Kenji

    2012-01-01

    The aim of the present study was to quantitatively evaluate the skin permeation/penetration of nanomaterials and to consider their penetration pathway through skin. Firstly, penetration/permeation of a model fluorescent nanoparticle, Fluoresbrite®, was determined through intact rat skin and several damaged skins. Fluoresbrite® permeated through only needle-punctured skin. The permeation profiles of soluble high molecular compounds, fluorescein isothiocyanate-dextrans (FITC-dextrans, FDs), with different molecular weights were also measured for comparison. The effects of molecular sizes and different skin pretreatments on the skin barrier were determined on the skin penetration/permeation of Fluoresbrite® and FDs. Fluoresbrite® was not permeated the intact skin, but FDs were permeated the skin. The skin distribution of titanium dioxide and zinc oxide nanoparticles was also observed after topical application of commercial cosmetics. Nanoparticles in sunscreen cosmetics were easily distributed into the groove and hair follicles after their topical application, but seldom migrated from the groove or follicles to viable epidermis and dermis. The obtained results suggested that nanoparticles did not permeate intact skin, but permeated pore-created skin. No or little permeation was observed for these nanomaterials through the stratum corneum.

  9. Lack of effect of selected sunscreens applied on ex vivo human skin for 5-methyl-aminolevulinic acid penetration and protoporphyrin IX photoactivation.

    Science.gov (United States)

    Osman-Ponchet, Hanan; Sevin, Karine; Gaborit, Alexandre; Kouidhi, Magali; Hanaizi, Johanna; Comby, Pierre; Ruty, Bernard; Bouvier, Guy

    2017-03-01

    Photodynamic therapy (PDT) is a successful treatment for non-melanoma skin cancers. Methyl-aminolevulinate (MAL) is metabolized to protoporphyrin IX (PpIX) which accumulates in the skin lesion and which generates a painful photochemical toxic reaction upon red light exposure. PDT using daylight (DL) exposure is now used to reduce pain and subjects are advised to protect the areas with sunscreen. This work investigated the effect of sunscreen on MAL penetration and PpIX photoactivation in ex vivo human skin. To measure skin penetration of MAL, particle-free sunscreens were applied on ex vivo human skin samples mounted on diffusion cells before application of Metvix cream containing [14C]-MAL for 2.5h. To circumvent the absence of skin penetration of PpIX, skin samples were first treated with microneedles and mounted on diffusion cells before the application of PpIX solution for 1h followed by sunscreens. Skin samples were then exposed to solar simulator for 1h. Concentrations of [14C]-MAL or PpIX were measured in both total skin and receptor liquid. The results showed that the in vitro skin penetration of MAL and the PpIX photoactivation on ex vivo human skin samples are not modified by pretreatments of ex vivo human skin with sunscreens. This study demonstrates that neither in vitro skin penetration of MAL nor PpIX photoactivation were modified by pretreatments with Cetaphil SPF 30 Dermacontrol and Actinica® Lotion SPF 50+. This supports the efficacy and safety of MAL DL-PDT in the clinical situation. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  10. Evaluation of skin permeation and analgesic activity effects of carbopol lornoxicam topical gels containing penetration enhancer.

    Science.gov (United States)

    Al-Suwayeh, Saleh A; Taha, Ehab I; Al-Qahtani, Fahad M; Ahmed, Mahrous O; Badran, Mohamed M

    2014-01-01

    The current study was designed to develop a topical gel formulation for improved skin penetration of lornoxicam (LOR) for enhancement of its analgesic activity. Moreover, the effect of different penetration enhancers on LOR was studied. The LOR gel formulations were prepared by using hydroxylpropyl methylcellulose (HPMC) and carbopol. The carbopol gels in presence of propylene glycol (PG) and ethanol were developed. The formulated gels were characterized for pH, viscosity, and LOR release using Franz diffusion cells. Also, in vitro skin permeation of LOR was conducted. The effect of hydroxypropyl β-cyclodextrin (HP β-CD), beta-cyclodextrin (β-CD), Tween 80, and oleic acid on LOR permeation was evaluated. The optimized LOR gel formulation (LORF8) showed the highest flux (14.31 μg/cm(2)/h) with ER of 18.34 when compared to LORF3. Incorporation of PG and HP β-CD in gel formulation (LORF8) enhanced the permeation of LOR significantly. It was observed that LORF3 and LORF8 show similar analgesic activity compared to marketed LOR injection (Xefo). This work shows that LOR can be formulated into carbopol gel in presence of PG and HP β-CD and may be promising in enhancing permeation.

  11. Evaluation of Skin Permeation and Analgesic Activity Effects of Carbopol Lornoxicam Topical Gels Containing Penetration Enhancer

    Directory of Open Access Journals (Sweden)

    Saleh A. Al-Suwayeh

    2014-01-01

    Full Text Available The current study was designed to develop a topical gel formulation for improved skin penetration of lornoxicam (LOR for enhancement of its analgesic activity. Moreover, the effect of different penetration enhancers on LOR was studied. The LOR gel formulations were prepared by using hydroxylpropyl methylcellulose (HPMC and carbopol. The carbopol gels in presence of propylene glycol (PG and ethanol were developed. The formulated gels were characterized for pH, viscosity, and LOR release using Franz diffusion cells. Also, in vitro skin permeation of LOR was conducted. The effect of hydroxypropyl β-cyclodextrin (HP β-CD, beta-cyclodextrin (β-CD, Tween 80, and oleic acid on LOR permeation was evaluated. The optimized LOR gel formulation (LORF8 showed the highest flux (14.31 μg/cm2/h with ER of 18.34 when compared to LORF3. Incorporation of PG and HP β-CD in gel formulation (LORF8 enhanced the permeation of LOR significantly. It was observed that LORF3 and LORF8 show similar analgesic activity compared to marketed LOR injection (Xefo. This work shows that LOR can be formulated into carbopol gel in presence of PG and HP β-CD and may be promising in enhancing permeation.

  12. The Effects of Geometry on Skin Penetration and Failure of Polymer Microneedles

    Science.gov (United States)

    Gittard, Shaun D; Chen, Bo; Xu, Huadong; Ovsianikov, Aleksandr; Chichkov, Boris N; Monteiro-Riviere, Nancy A; Narayan, Roger J

    2012-01-01

    Microneedles are small-scale devices that may be used for drug delivery and biosensing. In this study, the forces required for mechanical failure, the modes of mechanical failure, as well as the mechanisms for microneedle penetration into porcine skin were examined. Microneedles produced from the acrylate-based polymer e-Shell 200 using an indirect rapid prototyping approach involving two-photon polymerization and poly(dimethylsiloxane) micromolding were found to possess sufficient strength for penetration of porcine skin. The failure forces were an order of magnitude greater than the forces necessary for full insertion into the skin. Bending was the most common form of failure; an increasing aspect ratio and a decreasing tip diameter were associated with lower failure forces. Video captured during skin penetration revealed that microneedle penetration into the skin occurred by means of a series of insertions and not by means of a single insertion event. Images obtained during and after skin penetration confirmed microneedle penetration of skin as well as transdermal delivery of lucifer yellow dye. These findings shed insight into the mechanisms of microneedle penetration and failure, facilitating design improvements for polymer microneedles. PMID:23543070

  13. Fatty acids penetration into human skin ex vivo: A TOF-SIMS analysis approach.

    Science.gov (United States)

    Čižinauskas, Vytis; Elie, Nicolas; Brunelle, Alain; Briedis, Vitalis

    2017-03-02

    Linoleic, oleic, palmitoleic, palmitic, and stearic fatty acids (FAs) are commonly used in dermatological formulations. They differ by their structure, presence in the skin, and mode of application in pharmaceuticals and cosmetics compounding. These FAs are also known as chemical penetration enhancers, but their mechanisms of penetration enhancement and effect on barrier characteristics of the skin require additional study. In this study, the authors conducted an ex vivo analysis of the distribution of lipid components in the epidermis and dermis of human skin after applying individual FAs. The goal was to elucidate possible mechanisms of penetration enhancement and FA effects on barrier characteristics of the skin. FA penetration studies were conducted ex vivo on human skin and time-of-flight secondary ion mass spectrometry (TOF-SIMS) bioimaging analysis was performed to visualize and analyze distribution of FAs in skin sections. The current study demonstrated that TOF-SIMS imaging was effective in visualizing the distribution of linoleic, oleic, palmitoleic, palmitic, and stearic acid in the human skin ex vivo after the skin penetration experiment of individual FAs. The integration of the obtained TOF-SIMS images allowed a semiquantitative comparison of the effects induced by individual FA applications on the human skin ex vivo. FAs showed varying abilities to penetrate the skin and disorder the FAs within the skin, based on their structures and physicochemical properties. Linoleic acid penetrated the skin and changed the distribution of all the analyzed FAs. Skin treatment with palmitoleic or oleic acid increased the amounts of singular FAs in the skin. Penetration of saturated FAs was low, but it increased the detected amounts of linoleic acid in both skin layers. The results indicate that application of FAs on the skin surface induce redistribution of native FAs not only in the stratum corneum layer of epidermis but also in the lipid content of full epidermis

  14. In vitro penetration of alpha arbutin niosome span 60 system in gel preparation

    Directory of Open Access Journals (Sweden)

    Rise Desnita

    2017-12-01

    Full Text Available Alpha arbutin is a hydrophilic compound which is difficult to pass trough the stratum corneum. One of the effort to increase the penetration of the drug through the stratum corneum is by making in niosome system. This study aims to determine the optimum concentration of span 60 to improve the entrapment efficiency of niosome and investigate the increasing penetration of alpha arbutin using the niosome system in the preparation of the gel in vitro. Niosome consist a mixture of span 60 and cholesterol it made by thin layer hydration method. Concentration of span 60 was varied into three formulas were 100, 150 and 200 µmoL. The formulation of gel was made in two formulas including formulation of niosome alpha arbutin and alpha arbutin in gel. The determination of enterapment efficiency showed that formula 100µmol has an optimum enterapment efficiency by 99.09%±0.17.The in vitro penetration tests with shed snake skin membrane showed that usage span 60 as a niosome composer could increase penetration of alpha arbutin in gel formulation with cumulative numbers of diffusion in 8 hours was 91.62%±2.32 compared to alpha arbutin in gel without niosome system about 73.00%±0.94.

  15. The effect of skin aging on the percutaneous penetration of chemicals through human skin

    Energy Technology Data Exchange (ETDEWEB)

    Roskos, K.V.

    1989-01-01

    Despite much research into the mechanisms of cutaneous aging and the identification of significant age-associated biological and biophysical changes within the skin, the question how does aging affect percutaneous absorption (PA) in vivo remains unanswered. The author has made in vivo measurements of PA in young (18-40 years) and old (> 65 years) subjects. Standard radiotracer methodology was employed and PA was quantified from the urinary excretion profiles of {sup 14}C radiolabel (corrected for incomplete renal elimination). Testosterone (TST), estradiol (EST), hydrocortisone (HC), benzoic acid (BA), acetylsalicylic acid (ASA) and caffeine (CAFF) have been studied. Penetration of HC, BA, ASA, and CAFF were significantly lower in aged subjects whereas TST and EST absorption were not distinguishable from the young controls. Thus it appears that aging can affect PA in vivo and that relatively hydrophilic compounds may be most sensitive. Work was done to elucidate whether the observations were related to documented skin aging changes. Cutaneous microcirculation efficiency suspected to decline with increasing age, could not be correlated with the observed penetration changes. However, in vivo infrared spectroscopic studies of aged stratum corneum (SC) reveal a decreased amount of epidermal lipid. The diminished lipid content implies a diminished dissolution medium for compounds administered to the skin surface. They hypothesize that the compounds most affected by a loss of SC lipids would be those compounds whose overall solubility is lowest (compounds with lower octanol-water partition coefficients, eg., HC, BA, ASA and CAFF). Conversely, a diminished lipid content may not affect dissolution into the SC of highly lipophilic compounds (e.g., TST and EST).

  16. Psoralen loaded liposomal nanocarriers for improved skin penetration and efficacy of topical PUVA in psoriasis.

    Science.gov (United States)

    Doppalapudi, Sindhu; Jain, Anjali; Chopra, Dhiraj Kumar; Khan, Wahid

    2017-01-01

    Psoralen in combination with ultraviolet A radiation (PUVA) is an FDA recommended therapy for clinical application in the management of severe recalcitrant psoriasis. Psoralen acts by intercalation of DNA and upon exposure to UV-A, it forms monoadducts which in turn induce apoptosis. Poor skin deposition, weak percutaneous permeability of psoralen and adverse effects of severe burning, blisters, pigmentation associated with conventional topical psoralen vehicles hinders the therapeutic efficacy and safety of topical PUVA. The aim of the present study is to formulate psoralen loaded liposomal nanocarriers for enhanced skin penetration, safety and efficacy of topical PUVA in psoriasis. Two different liposomal compositions i.e., cationic liposomes composed of DC-Chol, cholesterol and anionic liposomes composed of egg lecithin, cholesterol, tetramyristoyl cardiolipin were prepared for the topical delivery of psoralen. Liposomal carriers were characterized with respect to size, zeta potential, entrapment efficiency, stability, in vitro drug release and in vivo studies. Both liposomes were prepared with particle size of nearly 100nm. Zeta potential and entrapment efficiency of cationic liposomes were +25.8mV, 75.12% and anionic liposomes were -28.5mV, 60.08% respectively. Liposomal dermal distribution demonstrated higher penetration of both liposomal carriers over solution. Similarly, skin permeation study indicated 5 fold increase in permeation of psoralen with liposomal carriers. Topical application of psoralen liposomal gels on imiquimod induced psoriatic plaque model reduced the symptoms of psoriasis and levels of key psoriatic cytokines such as tumor necrosis factor-α, IL-17 and IL-22. In conclusion, the developed liposomal carriers of psoralen were found to be promising and can find application for optimal safety and efficacy of topical PUVA in psoriasis. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Efficacy of skin wash on dermal absorption: an in vitro study on four model compounds of varying solubility

    DEFF Research Database (Denmark)

    Nielsen, Jesper Bo

    2010-01-01

    PURPOSE: Following dermal exposure to chemicals causing systemic toxicity, the general advice to avoid further systemic exposure is to wash the skin. The present study uses four model compounds (benzoic acid, glyphosat, caffeine, malathion) with varying size and solubility to substantiate...... this advice and quantify the effect of skin wash following 6 h dermal exposure on subsequent extent of skin penetration and deposition within the skin compartment. METHOD: Percutaneous penetration through human skin is studied in an in vitro model with static diffusion cells. RESULTS: The study demonstrates...

  18. In vitro percutaneous absorption in mouse skin: influence of skin appendages

    Energy Technology Data Exchange (ETDEWEB)

    Kao, J.; Hall, J.; Helman, G.

    1988-06-15

    Skin appendages are often envisaged as channels that bypass the stratum corneum barrier and are generally thought to facilitate the dermal absorption of topical agents. However, the significance of this transappendageal pathway in percutaneous absorption remains to be assessed experimentally. With the use of a skin organ culture penetration chamber system, the influence of skin appendages on the in vitro permeation of topically applied benzo(a)pyrene and testosterone (5 micrograms/2 cm2) was examined in skin preparations from both haired and hairless mice. Haired mice examined included the C57BL6, C3H, DBA2, Balbc, and Sencar strains and the hairless mice were the HRS and SKH. In all mouse strains examined, the overall permeation of testosterone (greater than 65% of applied dose) 16 hr following in vitro topical application was greater than that of benzo(a)pyrene (less than 10%). No strain differences were observed with respect to the percutaneous permeation of testosterone; however, percutaneous permeation of benzo(a)pyrene in the haired mice (7-10% of applied dose) was higher than that in the hairless mice (2%). In an in-house derived mouse strain which showed three phenotypic variants due to hair densities, the permeability to both compounds was highest in the skin of the haired phenotype (testosterone 67%, benzo(a)pyrene 7%), lowest in the hairless phenotype (35 and 1%, respectively) and intermediate in the fuzzy-haired animal (57 and 3%, respectively). Examination by fluorescence microscopy of cryosections of skin, prepared 1 hr after topical benzo(a)pyrene, showed areas of intense fluorescence deep within the nonfluorescing dermis of skin from the haired phenotype. These fluorescent areas were correlated with follicular ducts and sebaceous glands.

  19. Influence of massage and occlusion on the ex vivo skin penetration of rigid liposomes and invasomes

    DEFF Research Database (Denmark)

    Trauer, S.; Richter, H.; Kuntsche, Judith

    2014-01-01

    Liposomes are frequently described as drug delivery systems for dermal and transdermal applications. Recently, it has been shown that particulate substances penetrate effectively into hair follicles and that the follicular penetration depth can be increased by massaging the skin, which simulates...... the in vivo movement of hairs in the hair follicles. In the present study, massage was applied to skin mounted to Franz diffusion cells. By means of confocal laser scanning microscopy, the influence of massage and occlusion on the follicular penetration depths of rigid and flexible liposomes loaded...... with a hydrophilic and lipophilic dye was investigated. The application of massage increased follicular penetration significantly. Occlusion resulted in an increased follicular penetration depth only for rigid liposomes, whereas invasomes did not penetrate more effectively if occlusion was applied. The results...

  20. Quantification of skin penetration of antioxidants of varying lipophilicity.

    Science.gov (United States)

    Abla, M J; Banga, A K

    2013-02-01

    Antioxidants play a vital role in protecting the skin from environmental distress. As the skin is constantly exposed to harmful UV radiation, endogenous antioxidants present in the superficial layers of the skin neutralize reactive oxygen species. Over time, antioxidants become depleted and loss their protective effect on the skin. Therefore, supplementing skin with topical antioxidant can help replenish this loss and fight the oxidative stress. The objective of this study was to deliver antioxidants topically and quantify the amount permeated in the stratum corneum and underlying skin. Polyphenols (catechin, resveratrol and curcumin) and vitamin (retinol) with various lipophilic properties were delivered via porcine ear skin, using propylene glycol as a vehicle. The amount in the stratum corneum and underlying skin was quantified using tape stripping and skin extraction methods, respectively, and samples were analysed via HPLC. All four antioxidants permeated into the skin from the propylene glycol vehicle. The order of the amount of antioxidant in the stratum corneum was catechin > resveratrol~ retinol> curcumin, whereas that in the underlying skin was retinol > catechin~ resveratrol~ curcumin. Of the total amount of polyphenols in the skin, approximately 90% was retained in the stratum corneum whereas 10% was quantified in the underlying skin. In contrast, 10% of retinol was retained in the stratum corneum whereas 90% permeated in the underlying skin. Polyphenols (catechin, resveratrol and curcumin) showed high concentration in the stratum corneum whereas retinol showed high accumulation in the underlying layers of the skin. © 2012 The Authors ICS © 2012 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  1. Cationic membrane-active peptides - anticancer and antifungal activity as well as penetration into human skin.

    Science.gov (United States)

    Do, Nhung; Weindl, Günther; Grohmann, Lisa; Salwiczek, Mario; Koksch, Beate; Korting, Hans Christian; Schäfer-Korting, Monika

    2014-05-01

    Cationic antimicrobial peptides are ancient natural broad-spectrum antibiotics, and several compounds also exhibit anticancer activity. However, most applications pertain to bacterial infections, and treatment for skin cancer is less frequently considered. The cytotoxicity of melittin, cecropin A, protegrin-1 and histatin 5 against squamous skin cancer cell lines and normal human keratinocytes was evaluated and compared to established drugs. The results show that melittin clearly outperforms 5-fluorouracil regarding antitumor activity. Importantly, combined melittin and 5-fluorouracil enhanced cytotoxic effects on cancer cells and reduced toxicity on normal keratinocytes. Additionally, minimum inhibitory concentrations indicate that melittin also shows superior activity against clinical and laboratory strains of Candida albicans compared to amphotericin B. To evaluate its potential for topical applications, human skin penetration of melittin was investigated ex vivo and compared to two non-toxic cell-penetrating peptides (CPPs), low molecular weight protamine (LMWP) and penetratin. The stratum corneum prevents penetration into viable epidermis over 6 h; however, the peptides gain access to the viable skin after 24 h. Inhibition of digestive enzymes during skin penetration significantly enhances the availability of intact peptide. In conclusion, melittin may represent an innovative agent for non-melanoma skin cancer and infectious skin diseases. In order to develop a drug candidate, skin absorption and proteolytic digestion by skin enzymes need to be addressed. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  2. Validation of artificial skin equivalents as in vitro testing systems

    Science.gov (United States)

    Schmitt, Robert; Marx, Ulrich; Walles, Heike; Schober, Lena

    2011-03-01

    With the increasing complexity of the chemical composition of pharmaceuticals, cosmetics and everyday substances, the awareness of potential health issues and long term damages for humanoid organs is shifting into focus. Artificial in vitro testing systems play an important role in providing reliable test conditions and replacing precarious animal testing. Especially artificial skin equivalents ASEs are used for a broad spectrum of studies like penetration, irritation and corrosion of substances. One major challenge in tissue engineering is the qualification of each individual ASE as in vitro testing system. Due to biological fluctuations, the stratum corneum hornified layer of some ASEs may not fully develop or other defects might occur. For monitoring these effects we developed an fully automated Optical Coherence Tomography device. Here, we present different methods to characterize and evaluate the quality of the ASEs based on image and data processing of OCT B-scans. By analysing the surface structure, defects, like cuts or tears, are detectable. A further indicator for the quality of the ASE is the morphology of the tissue. This allows to determine if the skin model has reached the final growth state. We found, that OCT is a well suited technology for automatically characterizing artificial skin equivalents and validating the application as testing system.

  3. Influence of Characteristics of Oily Vehicle on Skin Penetration of Ufenamate.

    Science.gov (United States)

    Iino, Hayato; Fujii, Makiko; Fujino, Manami; Kohara, Shizuka; Hashizaki, Kaname; Kira, Hitomi; Koizumi, Naoya; Watanabe, Yoshiteru; Utoguchi, Naoki

    2017-01-01

    Skin penetration amounts of a highly lipophilic drug, ufenamate, prepared in four oily vehicles, including white petrolatum (WP), liquid paraffin (LP), isopropyl myristate (IPM), and isocetyl stearate (ICS), were compared. Ufenamate was mixed in each vehicle at 5% and applied at a rate of 2 mg/cm(2) to intact, stripped, and delipidized Yucatan micropig skin. The amounts of ufenamate and IPM in the stratum corneum (SC), epidermis, and dermis were determined. The skin penetration amounts of ufenamate from liquid oils were significantly higher than those from WP; the amounts of ufenamate were in the order WPskin penetration amount was approximately 20 times that of ufenamate. The skin penetration amounts of ufenamate from the liquid vehicles decreased after application to delipidized skin and were not significantly different among the four vehicles. The skin penetration amounts of the vehicle oils were significant and might disrupt intercellular lipid structures, especially in the strips 1-6 of the SC. In the deeper SC, there was no effect of the vehicle or skin condition. Thus, ufenamate mixed in liquid vehicles was found to be an effective dosage form.

  4. Size dependent skin penetration of nanoparticles in murine and porcine dermatitis models.

    Science.gov (United States)

    Try, Céline; Moulari, Brice; Béduneau, Arnaud; Fantini, Oscar; Pin, Didier; Pellequer, Yann; Lamprecht, Alf

    2016-03-01

    A major limitation in the current topical treatment of inflammatory skin diseases is the inability to selectively deliver the drug to the inflammation site. Recently, smart drug delivery systems such as nanocarriers are being investigated to enhance the selective deposition of anti-inflammatory drugs in inflamed areas of the skin to achieve higher therapeutic efficacy with minimal side effects. Of such systems, polymeric nanoparticles are considered very efficient carriers for the topical drug delivery. In the current work, poly(l-lactide-co-glycolide) nanoparticles of nominal sizes of 70nm (NP70) and 300nm (NP300) were studied for their intra-epidermal distribution in murine and pig atopic dermatitis models over time against the respective healthy controls. Confocal laser scanning microscopical examination of skin biopsies was utilized for the qualitative and semi-quantitative analyses of nanoparticles skin deposition and penetration depth. While no skin penetration was found for any of the particles in healthy skin, the accumulation of NP70 was significantly higher than NP300 in inflamed skin (15-fold in mice, 5-fold in pigs). Penetration depth of NP70 decreased over time in mice from 55±3μm to 20±2μm and similar tendencies were observed for the other formulations. In inflamed pig skin, a similar trend was found for the penetration depth (NP70: 46±12μm versus NP300: 23±3μm); however, the NP amount remained constant for the whole analyzed period. Their ability to penetrate specifically into inflamed skin combined with minimal effects on healthy skin underlines small polymeric nanoparticles' potential as selective drug carriers in future treatment of chronic inflammatory skin diseases such as atopic dermatitis. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Spatially Resolved Two-Color Diffusion Measurements in Human Skin Applied to Transdermal Liposome Penetration

    DEFF Research Database (Denmark)

    Brewer, Jonathan; Bloksgaard, Maria; Kubiak, Jakub

    2013-01-01

    of the liposome composition (phospholipids or transfersomes), our results show a clear lack of cross-correlation below the skin surface, indicating that the penetration of intact liposomes is highly compromised by the skin barrier.Journal of Investigative Dermatology advance online publication, 6 December 2012...

  6. In Vivo Human Skin Penetration Study of Sunscreens by Confocal Raman Spectroscopy.

    Science.gov (United States)

    Tippavajhala, Vamshi Krishna; de Oliveira Mendes, Thiago; Martin, Airton Abrahão

    2017-10-05

    This research work mainly deals with the application of confocal Raman spectroscopic technique to study in vivo human skin penetration of sunscreen products, as there are a lot of controversies associated with their skin penetration. Healthy human volunteers were tested for penetration of two commercial sunscreen products into their volar forearm skin for a period of 2 h. Measurements were taken before and after application of these sunscreen products. All the confocal Raman spectra were pre-processed and then subjected to multivariate two-dimensional principal component analysis and classical least squares analysis to determine the skin penetration of these sunscreens in comparison to the "sunscreen product spectrum" which was considered as the control. Score plots of principal component analysis of confocal Raman spectra indicated clear separation between the spectra before and after application of sunscreen products. Loading plots showed the maximum differences in the spectral region from 1590 to 1626 cm(-1) where the characteristic peak of the pure sunscreen products was observed. Classical least squares analysis has shown a significant penetration to a depth of 10 μm in the volar forearm skin of healthy human volunteers for both these sunscreen products. The results confirm that the penetration of these tested sunscreen products was restricted to stratum corneum and also prove that confocal Raman spectroscopy is a simple, fast, nondestructive, and noninvasive semi-quantitative analytical technique for these studies.

  7. Stimulation of the penetration of particles into the skin by plasma tissue interaction

    Science.gov (United States)

    Lademann, O.; Richter, H.; Kramer, A.; Patzelt, A.; Meinke, M. C.; Graf, C.; Gao, Q.; Korotianskiy, E.; Rühl, E.; Weltmann, K.-D.; Lademann, J.; Koch, S.

    2011-10-01

    A high number of treatments in dermatology are based on the penetration of topically applied drugs through the skin barrier. This process is predominantly inefficient, on account of the strong protection properties of the upper skin layer - the stratum corneum. If the skin barrier is damaged, the penetration efficiency of topically applied drugs increases. Therefore, different methods have been developed to influence the barrier properties of the skin. Recently, it could be demonstrated that a cold tissue tolerable plasma (TTP) produced by a plasma-jet can strongly enhance drug delivery through the skin. These investigations were performed by using a solution of fluorescent dye as a model drug. In the present study, these investigations were carried out using fluorescent silica particles at different sizes. The aim of the study was to investigate whether or not there is a limitation in size for topically applied substances to pass through the skin barrier after plasma treatment.

  8. Skin accumulation and penetration of a hydrophilic compound by a novel gemini surfactant, sodium dilauramidoglutamide lysine.

    Science.gov (United States)

    Hikima, Tomohiro; Tamura, Yoshinaga; Yamawaki, Yukio; Yamamoto, Masashi; Tojo, Kakuji

    2013-02-25

    We investigated a novel peptide-based gemini amphiphilic compound, sodium dilauramidoglutamide lysine (DLGL), as a chemical enhancer for the skin penetration of l-ascorbic acid 2-glucoside (AAG). A three-dimensional cultured human skin product, TESTSKIN™ LSE-high (LSE-high), was used as a skin model. The penetration flux of AAG with DLGL and that obtained with sodium lauramidoglutamide (LG) as a conventional surfactant across LSE-high were increased by 12.56 and 69.29 times compared to the control, respectively. The ratio of AAG amount with DLGL in the skin (21.78% total dose) was significantly increased (pskin, resulting in enhanced AAG penetration flux. However, LG might create the pathways through the skin. We conclude that DLGL is a gemini surfactant that accumulates a hydrophilic compound in skin and enhances the penetration flux. DLGL may therefore be a novel addition agent for skin local therapy. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. In vitro penetration of bleaching agents into the pulp chamber

    DEFF Research Database (Denmark)

    Benetti, Ana Raquel; Valera, M C; Mancini, M N G

    2004-01-01

    To investigate pulp chamber penetration of bleaching agents in teeth following restorative procedures.......To investigate pulp chamber penetration of bleaching agents in teeth following restorative procedures....

  10. Influence of massage and occlusion on the ex vivo skin penetration of rigid liposomes and invasomes.

    Science.gov (United States)

    Trauer, Sindy; Richter, Heike; Kuntsche, Judith; Büttemeyer, Rolf; Liebsch, Manfred; Linscheid, Michael; Fahr, Alfred; Schäfer-Korting, Monika; Lademann, Jürgen; Patzelt, Alexa

    2014-02-01

    Liposomes are frequently described as drug delivery systems for dermal and transdermal applications. Recently, it has been shown that particulate substances penetrate effectively into hair follicles and that the follicular penetration depth can be increased by massaging the skin, which simulates the in vivo movement of hairs in the hair follicles. In the present study, massage was applied to skin mounted to Franz diffusion cells. By means of confocal laser scanning microscopy, the influence of massage and occlusion on the follicular penetration depths of rigid and flexible liposomes loaded with a hydrophilic and lipophilic dye was investigated. The application of massage increased follicular penetration significantly. Occlusion resulted in an increased follicular penetration depth only for rigid liposomes, whereas invasomes did not penetrate more effectively if occlusion was applied. The results confirm that massage is an important tool for increasing follicular penetration in ex vivo studies using Franz diffusion cells. Occlusion may reduce the efficacy of follicular penetration depending on the specific liposomal preparation. Rigidity in particular appears to be a relevant parameter. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Effect of Size, Surface Charge, and Hydrophobicity of Poly(amidoamine) Dendrimers on Their Skin Penetration

    Science.gov (United States)

    Yang, Yang; Sunoqrot, Suhair; Stowell, Chelsea; Ji, Jingli; Lee, Chan-Woo; Kim, Jin Woong; Khan, Seema A.; Hong, Seungpyo

    2012-01-01

    The barrier functions of the stratum corneum (SC) and the epidermal layers present a tremendous challenge in achieving effective transdermal delivery of drug molecules. Although a few reports have shown that poly(amidoamine) (PAMAM) dendrimers are effective skin penetration enhancers, little is known regarding the fundamental mechanisms behind the dendrimer-skin interactions. In this paper, we have performed a systematic study to better elucidate how dendrimers interact with skin layers depending on their size and surface groups. Franz diffusion cells and confocal microscopy were employed to observe dendrimer interactions with full-thickness porcine skin samples. We have found that smaller PAMAM dendrimers (generation 2 (G2)) penetrate the skin layers more efficiently than the larger ones (G4). We have also found that G2 PAMAM dendrimers that are surface modified by either acetylation or carboxylation exhibit increased skin permeation and likely diffuse through an extracellular pathway. In contrast, amine-terminated dendrimers show enhanced cell internalization and skin retention but reduced skin permeation. In addition, conjugation of oleic acid (OA) to G2 dendrimers increases their 1-octanol/PBS partition coefficient, resulting in increased skin absorption and retention. Here we report that size, surface charge, and hydrophobicity directly dictate the permeation route and efficiency of dendrimer translocation across the skin layers, providing a design guideline for engineering PAMAM dendrimers as a potential transdermal delivery vector. PMID:22621160

  12. Penetration and decontamination of americium-241 ex vivo using fresh and frozen pig skin.

    Science.gov (United States)

    Tazrart, A; Bolzinger, M A; Moureau, A; Molina, T; Coudert, S; Angulo, J F; Briancon, S; Griffiths, N M

    2017-04-01

    Skin contamination is one of the most probable risks following major nuclear or radiological incidents. However, accidents involving skin contamination with radionuclides may occur in the nuclear industry, in research laboratories and in nuclear medicine departments. This work aims to measure the penetration of the radiological contaminant Americium ( 241 Am) in fresh and frozen skin and to evaluate the distribution of the contamination in the skin. Decontamination tests were performed using water, Fuller's earth and diethylene triamine pentaacetic acid (DTPA), which is the recommended treatment in case of skin contamination with actinides such as plutonium or americium. To assess these parameters, we used the Franz cell diffusion system with full-thickness skin obtained from pigs' ears, representative of human skin. Solutions of 241 Am were deposited on the skin samples. The radioactivity content in each compartment and skin layers was measured after 24 h by liquid scintillation counting and alpha spectrophotometry. The Am cutaneous penetration to the receiver compartment is almost negligible in fresh and frozen skin. Multiple washings with water and DTPA recovered about 90% of the initial activity. The rest remains fixed mainly in the stratum corneum. Traces of activity were detected within the epidermis and dermis which is fixed and not accessible to the decontamination. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  13. Comparison of the skin penetration of Garcinia mangostana extract in particulate and non-particulate form.

    Science.gov (United States)

    Tachaprutinun, Amornset; Meinke, Martina C; Richter, Heike; Pan-In, Porntip; Wanichwecharungruang, Supason; Knorr, Fanny; Lademann, Jürgen; Patzelt, Alexa

    2014-02-01

    The aim of the present study was to solve the water insolubility limitation of the medically and cosmetically interesting substance Garcinia mangostana Linn (GML) extract by encapsulation, and to evaluate and investigate the penetration efficacy of free and encapsulated GML in two different vehicles (water and cream) in porcine ear skin. The follicular penetration depth was determined in 50 hair follicles for each of the four formulations by means of fluorescence microscopy. Tape stripping was used to compare the distribution properties of GML with all formulations on the stratum corneum. The results showed that encapsulated and free GML in the cream base penetrated deeper into hair follicles than if applied in an aqueous base. In addition, encapsulated GML could be distributed more homogeneously on the stratum corneum than the free GML. In conclusion, it was found that encapsulated GML in a cream base had the most effective penetration level in porcine ear skin. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Evaluation of geometrical effects of microneedles on skin penetration by CT scan and finite element analysis.

    Science.gov (United States)

    Loizidou, Eriketi Z; Inoue, Nicholas T; Ashton-Barnett, Johnny; Barrow, David A; Allender, Chris J

    2016-10-01

    Computerized tomography scan (CT scan) imaging and finite element analysis were employed to investigate how the geometric composition of microneedles affects their mechanical strength and penetration characteristics. Simulations of microneedle arrays, comprising triangular, square and hexagonal microneedle base, revealed a linear dependence of the mechanical strength to the number of vertices in the polygon base. A laser-enabled, micromoulding technique was then used to fabricate 3×3 microneedle arrays, each individual microneedle having triangular, square or hexagonal base geometries. Their penetration characteristics into ex-vivo porcine skin, were investigated for the first time by CT scan imaging. This revealed greater penetration depths for the triangular and square-based microneedles, demonstrating CT scan as a powerful and reliable technique for studying microneedle skin penetration. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Microwave-aided skin drug penetration and retention of 5-fluorouracil-loaded ethosomes.

    Science.gov (United States)

    Khan, Nauman Rahim; Wong, Tin Wui

    2016-09-01

    Skin drug retention is required in local treatment of skin cancer. This study investigated the interplay effects of ethosomes and microwave in transdermal drug delivery. Skin pre-treatment by microwave and applied with liquified medicine is deemed to 'cement' the skin thereby raising skin drug deposition. 5-fluorouracil-loaded ethosomes were prepared and subjected to size, zeta potential, morphology, drug content, drug release and skin permeation tests. The molecular characteristics of untreated, microwave and/or ethosome-treated skins were examined by Fourier transform infrared and raman spectroscopy, thermal and electron microscopy techniques. The skin drug retention was promoted using larger ethosomes with negative zeta potentials that repelled anionic lipids of skin and hindered vesicle permeation into deep layers. These ethosomes had low ethanol content. They were less able to fluidize the lipid and defluidize the protein domains at epidermis to enlarge aqueous pores for drug permeation. Pre-treatment of skin by 2450 MHz microwave for 2.5 min further increased skin drug penetration and retention of low ethanol ethosomes and provided lower drug permeation than cases treated for 1.15 min and 5 min. A 2.5 min treatment might be accompanied by specific dermal protein fluidization via C=O moiety which translated to macromolecular swelling, narrowing of intercellular spaces at lower skin layers, increased drug retention and reduced drug permeation. Ethosomes and microwave synergized to promote skin drug retention.

  16. Penetration of topically applied nanocarriers into the hair follicles of dog and rat dorsal skin and porcine ear skin.

    Science.gov (United States)

    Knorr, Fanny; Patzelt, Alexa; Darvin, Maxim E; Lehr, Claus-Michael; Schäfer, Ulrich; Gruber, Achim D; Ostrowski, Anja; Lademann, Jürgen

    2016-08-01

    In humans, topically applied nanocarriers penetrate effectively into the hair follicles where they can be exploited for the localized and targeted treatment of skin disorders. The objective of the present study was to examine the applicability of particle-based systems for follicular drug delivery in companion animals and livestock, which have a large follicular reservoir. Skin samples from 10 beagle dogs, 14 Wistar rats and four ears from freshly slaughtered cross-bred pigs were used. Fluoresceinamine labelled poly (L-lactide-co-glycolide) nanocarriers (256 or 430 nm) were applied on the different skin samples. After penetration, skin biopsies were removed and cryohistological cross sections prepared and investigated with regard to the follicular penetration depths (in μm ± standard deviation) of the nanocarriers using confocal laser scanning microscopy. In canine, rat and porcine hair follicles, the smaller nanoparticles were detected at mean follicular penetration depths of 630.16 ± 135.75 μm, 253.55 ± 47.36 μm and 653.40 ± 94.71 μm, respectively. The larger particles were observed at average follicular depths of 604.79 ± 132.42 μm; 262.87 ± 55.25 μm and 786.81 ± 121.73 μm, respectively, in canine, rat and porcine hair follicles. Statistically significant differences (P porcine skin samples. The mean follicular penetration depths of the differently sized nanocarriers were mostly significantly different between the different species, which might be due to different species-specific follicular dimensions. This issue needs to be addressed specifically in further studies. © 2016 ESVD and ACVD.

  17. In vivo investigations on the penetration of various oils and their influence on the skin barrier.

    Science.gov (United States)

    Patzelt, A; Lademann, J; Richter, H; Darvin, M E; Schanzer, S; Thiede, G; Sterry, W; Vergou, T; Hauser, M

    2012-08-01

    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. 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. 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. 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. © 2011 John Wiley & Sons A/S.

  18. Effect of skin tumor properties on laser penetration

    CSIR Research Space (South Africa)

    Karsten, AE

    2009-06-01

    Full Text Available , data from literature can be used. One of the *AKarsten@csir.co.za; phone +(27)12 841 3428; fax +(27)12 841 3152 Therapeutic Laser Applications and Laser-Tissue Interactions IV, edited by Ronald Sroka, Lothar D. Lilge, Proc. of SPIE-OSA Biomedical...]. Three different non-melanoma skin cancers: nodular basal cell carcinoma (NBCC), infiltrative basal cell carcinoma (IBCC) and squamous cell carcinoma (SCC) were investigated. In the “control” model, the properties of the tumor were kept the same...

  19. Fiber optic microneedles for transdermal light delivery: ex vivo porcine skin penetration experiments.

    Science.gov (United States)

    Kosoglu, Mehmet A; Hood, Robert L; Chen, Ye; Xu, Yong; Rylander, Marissa Nichole; Rylander, Christopher G

    2010-09-01

    Shallow light penetration in tissue has been a technical barrier to the development of light-based methods for in vivo diagnosis and treatment of epithelial carcinomas. This problem can potentially be solved by utilizing minimally invasive probes to deliver light directly to target areas. To develop this solution, fiber optic microneedles capable of delivering light for either imaging or therapy were manufactured by tapering step-index silica-based optical fibers employing a melt-drawing process. Some of the microneedles were manufactured to have sharper tips by changing the heat source during the melt-drawing process. All of the microneedles were individually inserted into ex vivo pig skin samples to demonstrate the feasibility of their application in human tissues. The force on each microneedle was measured during insertion in order to determine the effects of sharper tips on the peak force and the steadiness of the increase in force. Skin penetration experiments showed that sharp fiber optic microneedles that are 3 mm long penetrate through 2 mm of ex vivo pig skin specimens. These sharp microneedles had a minimum average diameter of 73 mum and a maximum tip diameter of 8 mum. Flat microneedles, which had larger tip diameters, required a minimum average diameter of 125 mum in order to penetrate through pig skin samples. Force versus displacement plots showed that a sharp tip on a fiber optic microneedle decreased the skin's resistance during insertion. Also, the force acting on a sharp microneedle increased more steadily compared with a microneedle with a flat tip. However, many of the sharp microneedles sustained damage during skin penetration. Two designs that did not accrue damage were identified and will provide a basis of more robust microneedles. Developing resilient microneedles with smaller diameters will lead to transformative, novel modes of transdermal imaging and treatment that are less invasive and less painful for the patient.

  20. Skin penetration and retention of L-ascorbic acid 2-phosphate using multilamellar vesicles.

    Science.gov (United States)

    Yoo, Juno; Shanmugam, Srinivasan; Song, Chung-Kil; Kim, Dae-Duk; Choi, Han-Gon; Yong, Chul-Soon; Woo, Jong-Soo; Yoo, Bong Kyu

    2008-12-01

    Transdermal formulation of L-ascorbic acid 2-phosphate magnesium salt (A2P) was prepared using multilamellar vesicles (MLV). A2P was either physically mixed with or entrapped into three different MLVs of neutral, cationic, and anionic liposome vesicles. For the preparation of neutral MLVs, phosphatidylcholine (PC) and cholesterol (CH) were used. For cationic and anionic MLVs, dioleoyl-trimethylammonium-propane and dimyristoyl glycerophosphate were added as surface charge inducers, respectively, in addition to PC and CH. Particle size of the three A2P-loaded MLVs was submicron, and polydispersity index revealed homogenous distribution of the prepared MLVs except neutral ones. Skin penetration study with hairless mouse skin showed that both physical mixtures of A2P with empty MLVs and A2P-loaded MLVs increased penetration of the drug compared to aqueous A2P solution. During the penetration, however, significant amount of the drug was metabolized into L-ascorbic acid, which has no beneficial effect on stimulation of hair growth. Out of the physical mixtures and A2P-loaded MLVs tested, physical mixture of A2P with empty cationic MLV resulted in the greatest skin penetration and retention in hairless mouse skin.

  1. Human skin penetration and local effects of topical nano zinc oxide after occlusion and barrier impairment.

    Science.gov (United States)

    Leite-Silva, V R; Sanchez, W Y; Studier, H; Liu, D C; Mohammed, Y H; Holmes, A M; Ryan, E M; Haridass, I N; Chandrasekaran, N C; Becker, W; Grice, J E; Benson, H A E; Roberts, M S

    2016-07-01

    Public health concerns continue to exist over the safety of zinc oxide nanoparticles that are commonly used in sunscreen formulations. In this work, we assessed the effects of two conditions which may be encountered in everyday sunscreen use, occlusion and a compromised skin barrier, on the penetration and local toxicity of two topically applied zinc oxide nanoparticle products. Caprylic/capric triglyceride (CCT) suspensions of commercially used zinc oxide nanoparticles, either uncoated or with a silane coating, were applied to intact and barrier impaired skin of volunteers, without and with occlusion for a period of six hours. The exposure time was chosen to simulate normal in-use conditions. Multiphoton tomography with fluorescence lifetime imaging was used to noninvasively assess zinc oxide penetration and cellular metabolic changes that could be indicative of toxicity. We found that zinc oxide nanoparticles did not penetrate into the viable epidermis of intact or barrier impaired skin of volunteers, without or with occlusion. We also observed no apparent toxicity in the viable epidermis below the application sites. These findings were validated by ex vivo human skin studies in which zinc penetration was assessed by multiphoton tomography with fluorescence lifetime imaging as well as Zinpyr-1 staining and toxicity was assessed by MTS assays in zinc oxide treated skin cryosections. In conclusion, applications of zinc oxide nanoparticles under occlusive in-use conditions to volunteers are not associated with any measurable zinc oxide penetration into, or local toxicity in the viable epidermis below the application site. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Skin penetration behaviour of liposomes as a function of their composition.

    Science.gov (United States)

    Gillet, A; Lecomte, F; Hubert, P; Ducat, E; Evrard, B; Piel, G

    2011-09-01

    Deformable liposomes have been developed and evaluated as a novel topical and transdermal delivery system. Their mechanism of drug transport into and through the skin has been investigated but remains a much debated question. The present study concerns ex vivo diffusion experiments using pig ear skin in order to explain the penetration mechanism of classical and deformable liposomes. Classical and deformable vesicles containing betamethasone in the aqueous compartment through the use of cyclodextrin inclusion complexes were compared to vesicles encapsulating betamethasone in their lipid bilayer. Deformable liposomes contained sodium deoxycholate as the edge activator. Liposomes were characterised by their diameter, encapsulation efficiency, deformability, stability (in terms of change in diameter) and release of encapsulated drug. Ex vivo diffusion studies using Franz diffusion cells were performed. Confocal microscopy was performed to visualise the penetration of fluorescently labelled liposomes into the skin. This study showed that liposomes do not stay intact when they penetrate the deepest layers of the skin. Betamethasone is released from the vesicles after which free drug molecules can diffuse through the stratum corneum and partition into the viable skin tissue. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Skin tissue engineering--in vivo and in vitro applications.

    Science.gov (United States)

    Groeber, Florian; Holeiter, Monika; Hampel, Martina; Hinderer, Svenja; Schenke-Layland, Katja

    2011-04-30

    Significant progress has been made over the years in the development of in vitro-engineered substitutes that mimic human skin, either to be used as grafts for the replacement of lost skin or for the establishment of human-based in vitro skin models. This review summarizes these advances in in vivo and in vitro applications of tissue-engineered skin. We further highlight novel efforts in the design of complex disease-in-a-dish models for studies ranging from disease etiology to drug development and screening. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Photoacoustic evaluation of the penetration of piroxicam gel applied with phonophoresis into human skin

    Energy Technology Data Exchange (ETDEWEB)

    Silveira, F L F D; Barja, P R [Research and Development Institute, UNIVAP, Av. Shishima Hifumi 2911, Sao Jose dos Campos, SP, 12209-010 (Brazil); Acosta-Avalos, D, E-mail: barja@univap.b [Centro Brasileiro de Pesquisas Fisicas (CBPF), R.Xavier Sigaud 150, Rio de Janeiro, RJ, 22290-180 (Brazil)

    2010-03-01

    The photoacoustic (PA) technique has been increasingly employed in biomedical studies, allowing in vivo skin measurements not easily performed with other techniques. It is possible to use PA measurements to evaluate transdermal delivery of products topically applied through manual massage or phonophoresis, that is the utilization of ultrasound waves to enhance drug absorption. The aim of this study was to analyze the influence of the period of phonophoresis application in the transdermal penetration of piroxicam gel. In vivo PA measurements employed a tungsten lamp as light source and a thin aluminum foil closing the PA chamber. The PA signals of the arm (i) clean; and (ii) after phonophoresis were utilized to estimate the concentration of piroxicam into skin. For all (4) volunteers, drug concentration in skin after phonophoresis application was the same for the different application times employed; in this way, phonophoresis for one minute seemed to be sufficient to enhance piroxicam penetration into skin. The actual amount of drug delivered into tissue depends on the person, suggesting a dependency with the skin type, which affects the PA signal level [2]. We conclude that drug delivery depends not only on the application method, but also on the specific skin type.

  5. Is skin penetration a determining factor in skin sensitization potential and potency? Refuting the notion of a LogKow threshold for Skin Sensitization

    Science.gov (United States)

    Summary:Background. It is widely accepted that substances that cannot penetrate through the skin will not be sensitisers. Thresholds based on relevant physicochemical parameters such as a LogKow > 1 and a MW < 500, are assumed and widely accepted as self-evident truths. Objective...

  6. Penetration of varnishes into demineralized root dentine in vitro

    NARCIS (Netherlands)

    Arends, J; Duschner, H; Ruben, JL

    1997-01-01

    In this paper the penetration of three different varnishes employed in caries prevention (Duraphat(R), Fluor Protector(R) and Cervitec(R)) into demineralized dentine is quantified using confocal laser scanning microscopy. The results show that the varnish penetration into lesions about 85 mu m in

  7. In vitro percutaneous penetration and characterization of silver from silver-containing textiles.

    Science.gov (United States)

    Bianco, Carlotta; Kezic, Sanja; Crosera, Matteo; Svetličić, Vesna; Šegota, Suzana; Maina, Giovanni; Romano, Canzio; Larese, Francesca; Adami, Gianpiero

    2015-01-01

    The objective of this study was to determine the in vitro percutaneous penetration of silver and characterize the silver species released from textiles in different layers of full thickness human skin. For this purpose, two different wound dressings and a garment soaked in artificial sweat were placed in the donor compartments of Franz cells for 24 hours. The concentration of silver in the donor phase and in the skin was determined by an electrothermal atomic absorption spectrometer (ET-AAS) and by inductively coupled plasma mass spectrometer (ICP-MS). The characterization of silver species in the textiles and in the skin layers was made by scanning electron microscopy with integrated energy dispersive X-ray spectroscopy (SEM-EDX). Additionally, the size distribution of silver nanoparticles in the textiles was performed by atomic force microscopy (AFM). On the surface of all investigated materials, silver nanoparticles of different size and morphology were found. Released silver concentrations in the soaking solutions (ie, exposure concentration) ranged from 0.7 to 4.7 μg/mL (0.6-4.0 μg/cm(2)), fitting the bactericidal range. Silver and silver chloride aggregates at sizes of up to 1 μm were identified both in the epidermis and dermis. The large size of these particles suggests that the aggregation occurred in the skin. The formation of these aggregates likely slowed down the systemic absorption of silver. Conversely, these aggregates may form a reservoir enabling prolonged release of silver ions, which might lead to local effects.

  8. Determination of three ultraviolet filters in sunscreen formulations and from skin penetration studies by high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Fernanda Maria Pinto Vilela

    2011-01-01

    Full Text Available An analytical procedure to quantify 3-benzophenone, octylmethoxycinnamate and octylsalicylate was validated and employed to assess these ultraviolet filters in sunscreen formulations and from skin penetration studies. The effect of the vehicle on the skin retention of these filters was investigated. HPLC and extraction procedure were found to be reliable when obtaining data for the sunscreen formulations and for evaluation skin penetration. The results demonstrated that a cream gel generated higher epidermal concentrations of these filters than a lotion or cream-based formulation. Additionally, when comparing the skin retentions of each filter using the same formulation, 3-benzophenone showed the highest skin retention.

  9. Efficacy of skin wash on dermal absorption: an in vitro study on four model compounds of varying solubility.

    Science.gov (United States)

    Nielsen, Jesper Bo

    2010-08-01

    Following dermal exposure to chemicals causing systemic toxicity, the general advice to avoid further systemic exposure is to wash the skin. The present study uses four model compounds (benzoic acid, glyphosat, caffeine, malathion) with varying size and solubility to substantiate this advice and quantify the effect of skin wash following 6 h dermal exposure on subsequent extent of skin penetration and deposition within the skin compartment. Percutaneous penetration through human skin is studied in an in vitro model with static diffusion cells. The study demonstrates that percutaneous penetration continues after end of exposure due to the reservoir present in the skin. However, penetration rate will decrease significantly, and it is evident that simple hand-wash after end of exposure not only reduces the amount of residue present in the upper skin compartment but also significantly reduces the total absorption of test substance, most so for the hydrophilic compounds. Our observations support the continuing initiatives from occupational hygienists to urge people with dermal exposure hazards to wash potentially exposed skin areas.

  10. Rutin—Increased Antioxidant Activity and Skin Penetration by Nanocrystal Technology (smartCrystals

    Directory of Open Access Journals (Sweden)

    Sung Min Pyo

    2016-02-01

    Full Text Available Rutin is a well-known antioxidant from the group of flavonoids. Its use in cosmetic dermal products is, however, limited due to its poor water solubility. In order to increase rutin saturation solubility and improve the diffusion to the skin, rutin nanocrystals were produced by the smartCrystal process, e.g., bead milling followed by high pressure homogenization. Rutin nanocrystals were further incorporated into hydroxypropyl cellulose (HPC gel and its long-term stability was assessed. Determination of the antioxidant activity was made by the DPPH (2,2-diphenyl-1-picrylhydrazyl assay for these formulations: rutin nanocrystals (mean size 300 nm, rutin raw drug powder (mean size 33 μm and commercial product. Furthermore, the skin penetration profile of rutin was investigated by the tape-stripping method on porcine skin. This study demonstrated that rutin nanocrystal gel had the highest neutralizing activity (90%, followed by a commercial product and rutin raw drug powder. According to the skin study, rutin nanocrystals penetrated to the deeper layers of the stratum corneum, the horny layer of the skin.

  11. Tuning the Extent and Depth of Penetration of Flexible Liposomes in Human Skin.

    Science.gov (United States)

    Franzé, Silvia; Donadoni, Giulia; Podestà, Alessandro; Procacci, Patrizia; Orioli, Marica; Carini, Marina; Minghetti, Paola; Cilurzo, Francesco

    2017-06-05

    In this work we made an attempt to assess the effect of drug-induced changes of flexibility on the penetration of deformable vesicles into the human skin. Eight cationic liposomes with different degrees of flexibility were obtained by entrapping unfractionated heparin, enoxaparin, and nadroparin. The deformability was studied by a novel, facile, and reliable extrusion assay appositely developed and validated by means of quantitative nanoscale mechanical AFM measurements of vesicle elastic modulus (log10(YM)). The proposed extrusion assay, determining the forces involved in vesicles deformation, resulted very sensitive to evidence of minimal changes in bilayer rigidity (σ) and vesicle deformation (K). The drug loading caused a reduction of liposome flexibility with respect to the reference plain liposomes and in accordance to the heparin type, drug to cationic lipid (DOTAP) ratio, and drug distribution within the vesicles. Interestingly, the σ and log10(YM) values perfectly correlated (R(2) = 0.935), demonstrating the reliability of the deformability data obtained with both approaches. The combination of TEM and LC-MS/MS spectrometry allowed the pattern of the penetration of the entire vesicles into the skin to be followed. In all cases, intact liposomes in the epidermis layers were observed and a relationship between the depth of penetration and the liposome flexibility was found, supporting the hypothesis of the whole vesicle penetration mechanism. Moreover, the results of the extent (R24) of vesicle penetration in the human skin samples showed a direct relation to the flexibility values (σ1 = 0.65 ± 0.10 MPa → R24 = 3.33 ± 0.02 μg/mg; σ2 = 0.95 ± 0.04 MPa → R24 = 1.18 ± 0.26 μg/mg; σ3 = 1.89 ± 0.30 MPa → R24 = 0.53 ± 0.33 μg/mg).

  12. Evaluation of Zona Pellucida Function for Sperm Penetration During In Vitro Fertilization in Pigs

    Science.gov (United States)

    TANIHARA, Fuminori; NAKAI, Michiko; KANEKO, Hiroyuki; NOGUCHI, Junko; OTOI, Takeshige; KIKUCHI, Kazuhiro

    2013-01-01

    Abstract In porcine oocytes, the function of the zona pellucida (ZP) with regard to sperm penetration or prevention of polyspermy is not well understood. In the present study, we investigated the effects of the ZP on sperm penetration during in vitro fertilization (IVF). We collected in vitro-matured oocytes with a first polar body (ZP+ oocytes). Some of them were freed from the ZP (ZP− oocytes) by two treatments (pronase and mechanical pipetting), and the effects of these treatments on sperm penetration parameters (sperm penetration rate and numbers of penetrated sperm per oocyte) were evaluated. There was no evident difference in the parameters between the two groups. Secondly, we compared the sperm penetration parameters of ZP+ and ZP− oocytes using frozen-thawed epididymal spermatozoa from four boars. Sperm penetration into ZP+ oocytes was found to be accelerated relative to ZP− oocytes. Thirdly, we evaluated the sperm penetration of ZP+ and ZP− oocytes at 1−10 h after IVF (3 h gamete co-incubation). The proportions of oocytes penetrated by sperm increased significantly with time in both groups; however, the number of penetrated sperm per oocyte did not increase in ZP− oocytes. Finally, we performed IVF using ZP− oocytes divided into control (3 h) and prolonged gamete co-incubation (5 h) groups. Greater numbers of sperm penetrated in the 5 h group than in the control group. These results suggest that the ZP and oolemma are not competent factors for prevention of polyspermy in our present porcine IVF system. However, it appears that ZP removal is one of the possibilities for reducing polyspermic penetration in vitro in pigs. PMID:23666494

  13. In vitro permeation and disposition of niacinamide in silicone and porcine skin of skin barrier-mimetic formulations.

    Science.gov (United States)

    Haque, Tasnuva; Lane, Majella E; Sil, Bruno C; Crowther, Jonathan M; Moore, David J

    2017-03-30

    Niacinamide (NIA) is an amide form of vitamin B3 which is used in cosmetic formulations to improve various skin conditions and it has also been shown to increase stratum corneum thickness following repeated application. In this study, three doses (5, 20 and 50μL per cm 2 ) of two NIA containing oil-in-water skin barrier-mimetic formulations were evaluated in silicone membrane and porcine ear skin and compared with a commercial control formulation. Permeation studies were conducted over 24h in Franz cells and at the end of the experiment membranes were washed and niacinamide was extracted. For the three doses, retention or deposition of NIA was generally higher in porcine skin compared with silicone membrane, consistent with the hydrophilic nature of the active. Despite the control containing a higher amount of active, comparable amounts of NIA were deposited in skin for all formulations for all doses; total skin absorption values (permeation and retention) of NIA were also comparable across all formulations. For infinite (50μL) and finite (5μL) doses the absolute permeation of NIA from the control formulation was significantly higher in porcine skin compared with both test formulations. This likely reflects differences in formulation components and/or presence of skin penetration enhancers in the formulations. Higher permeation for the 50 and 20μL dose was also evident in porcine skin compared with silicone membrane but the opposite is the case for the finite dose. The findings point to the critical importance of dose and occlusion when evaluating topical formulations in vitro and also the likelihood of exaggerated effects of excipients on permeation at infinite and pseudo-finite dose applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Photoacoustic Analysis of the Penetration Kinetics of Cordia verbenacea DC in Human Skin

    Science.gov (United States)

    Carvalho, S. S.; Barja, P. R.

    2012-11-01

    Phonophoresis consists of the utilization of ultrasound radiation associated to pharmacological agents in order to enhance transdermal penetration of applied drugs. It is a widely employed resource in physiotherapy practice, normally associated with anti-inflammatory drugs, such as Acheflan. This drug was developed in Brazil from the essential oil of Cordia verbenacea DC, a native plant of the Brazilian southern coast. In previous studies, the photoacoustic (PA) technique proved effective in the study of the penetration kinetics of topically applied products and in the evaluation of drug delivery after phonophoresis application. The present work aimed to evaluate the penetration kinetics of Acheflan in human skin, employing in vivo PA measurements after massage application or phonophoresis application. Ten volunteers (aged between 18 and 30 years) took part in the study. Time evolution of the PA signal was fitted to a Boltzmann curve, S-shaped. After statistical analysis, PA measurements have shown drug penetration for both application forms, but drug delivery was more evident after phonophoresis application, with a characteristic penetration time of less than 15 min for the stratum corneum.

  15. In vitro-in vivo correlation in skin permeation.

    Science.gov (United States)

    Mohammed, D; Matts, P J; Hadgraft, J; Lane, M E

    2014-02-01

    In vitro skin permeation studies have been used extensively in the development and optimisation of delivery of actives in vivo. However, there are few reported correlations of such in vitro studies with in vivo data. The aim of this study was to investigate the skin permeation of a model active, niacinamide, both in vitro and in vivo. Conventional diffusion cell studies were conducted in human skin to determine niacinamide permeation from a range of vehicles which included dimethyl isosorbide (DMI), propylene glycol (PG), propylene glycol monolaurate (PGML), N-methyl 2-pyrrolidone (NMP), Miglyol 812N® (MG), and mineral oil (MO). Single, binary or ternary systems were examined. The same vehicles were subsequently examined to investigate niacinamide delivery in vivo. For this proof-of-concept study one donor was used for the in vitro studies and one volunteer for the in vivo investigations to minimise biovariability. Analysis of in vitro samples was conducted using HPLC and in vivo uptake of niacinamide was evaluated using Confocal Raman spectroscopy (CRS). The amount of niacinamide permeated through skin in vitro was linearly proportional to the intensity of the niacinamide signal determined in the stratum corneum in vivo. A good correlation was observed between the signal intensities of selected vehicles and niacinamide signal intensity. The findings provide further support for the use of CRS to monitor drug delivery into and across the skin. In addition, the results highlight the critical role of the vehicle and its disposition in skin for effective dermal delivery.

  16. Penetration of tamoxifen citrate loaded ethosomes and liposomes across human skin: a comparative study with confocal laser scanning microscopy.

    Science.gov (United States)

    Sarwa, Khomendra K; Suresh, Preeti K; Rudrapal, Mithun; Verma, Vinod K

    2014-01-01

    In the present study, ethosomal and liposomal formulations containing tamoxifen citrate were prepared and evaluated for their penetration properties in human cadaver skin using Franz diffusion cell and confocal laser scanning microscope (CLSM). The results clearly revealed that ethosomal vesicles showed a better drug permeation profile than that of liposomal vesicles. In addition, low fluorescence intensity in CLSM was recorded with liposomes as compared to ethosomes, indicating lower cumulative amount of drug permeation from liposomal vesicles. Furthermore, CLSM showed uniform fluorescence intensity across the entire depth of skin in ethosomal treatment, indicating high penetrability of ethosomal vesicles through human cadaver skin. In contrast, low penetrability of conventional liposomal vesicles was recorded as penetration was limited to the 7(th) section (i.e. upper epidermis layer) of skin as evident from visualization of intact liposomal vesicles in CLSM.

  17. In vitro and human testing strategies for skin irritation.

    Science.gov (United States)

    Robinson, M K; Osborne, R; Perkins, M A

    2000-01-01

    Prior to the manufacture, transport, and marketing of chemicals or products, it is critical to assess their potential for skin toxicity (corrosion or irritation), thereby protecting the worker and consumer from adverse skin effects due to intended or accidental skin exposure. Traditionally, animal testing procedures have provided the data needed to assess the more severe forms of skin toxicity, and current regulations may require animal test data before permission can be obtained to manufacture, transport, or market chemicals or the products that contain them. In recent years, the use of animals to assess skin safety has been opposed by some as inhumane and unnecessary. The conflicting needs of the industrial toxicologist to (1) protect human safety, (2) comply with regulations, and (3) reduce animal testing have led to major efforts to develop alternative, yet predictive, test methods. A variety of in vitro skin corrosion test methods have been developed and several have successfully passed initial international validation. These have included skin or epidermal equivalent assays that have been shown to distinguish corrosive from noncorrosive chemicals. These skin/epidermal equivalent assays have also been modified and used to assess skin irritation potential relative to existing human exposure test data. The data show a good correlation between in vitro assay data and different types of human skin irritation data for both chemicals and consumer products. The effort to eliminate animal tests has also led to the development of a novel human patch test for assessment of acute skin irritation potential. A case study shows the benefits of in vitro and human skin irritation tests compared to the animal tests they seek to replace, and strategies now exist to adequately assess human skin irritation potential without the need to rely on animal test methods.

  18. Sperm penetration through the human zona pellucida as a predictor of in vitro fertilization.

    Science.gov (United States)

    Morales, P; Vantman, D; Madariaga, M

    1999-05-01

    The aim of this study was to determine the predictive value of sperm penetration into the perivitelline space of human cadaveric oocytes on in vitro fertilization outcome. Forty-two patients with tubal infertility undergoing ovarian stimulation with gonadotropin for in vitro fertilization and embryo transfer participated in the study. The number of spermatozoa bound to the human zona pellucida, the percentage of cadaveric oocytes with one or more spermatozoa in the perivitelline space, and the in vitro fertilization outcome were evaluated. Spermatozoa from 37 of 42 patients were able to penetrate the perivitelline space of cadaveric oocytes as well as to fertilize human oocytes in vitro. In three individuals, no penetration of the perivitelline space of cadaveric oocytes was observed and no in vitro fertilization was detected. Only two patients were able to fertilize the couple's oocytes without penetration of the cadaveric oocytes. Based on these results the specificity and the sensitivity of the assay to predict in vitro fertilization was 100% and 94.1%, respectively. Accordingly, these results suggest that sperm-zona penetration is a useful bioassay to predict male fertility potential in IVF outcome.

  19. In Vitro and In Vivo Measurement of Percutaneous Penetration of Low Molecular Weight Toxins of Military Interest

    Science.gov (United States)

    1990-10-21

    acetylsalicylic acid and urea (Bronaugh et al., 1982). However, it has been proposed that partition coefficients near one favor skin penetration by allowing the...Weanling pig skin was used as a model for human skin because it has similar permeability properties (3). EXPERIMENTAL METHODS MATERIALS Unlabeled PbTx-3...barrier properties of these two skin types were found to be similar (results not shown). In vivo percutaneous penetration of PbTx-3 This study was

  20. A Method for Quantification of Penetration of Nanoparticles through Skin Layers Using Near-Infrared Optical Imaging

    Directory of Open Access Journals (Sweden)

    Melinda Stees

    2015-07-01

    Full Text Available Our study presents a new method for tracking nanoparticle penetration through different layers of the skin using near-infrared dye-loaded nanoparticles (hydrodynamic diameter = 156 nm and optical imaging. The dye-loaded nanoparticles were mixed in a topical skin cream, applied to human cadaver skin and incubated either for three or 24 h post-application, skin tissue was clipped between glass slides prior to imaging for signal intensity across the skin thickness using an optical imaging system. The data show that nanoparticles penetrate through all the layers of the skin but there is almost an exponential decay in the signal intensity from epidermis to dermis. Depending upon the incubation time, about 55%–59% of the total signal was seen in the epidermis and the remaining through dermis and hypodermis. The advantage of the method is that it allows quantitative analysis of the extent of penetration of nanoparticles through different layers of the skin without interference of any background signal from skin tissue, and without requiring extensive tissue processing. Our method could potentially be used to study the effect of nanoparticle properties and/or the use of different formulation additives on penetration of nanoparticles through different skin layers.

  1. Loss of surface coat by Strongyloides ratti infective larvae during skin penetration: evidence using larvae radiolabelled with /sup 67/gallium

    Energy Technology Data Exchange (ETDEWEB)

    Grove, D.I.; Northern, C.; Warwick, A.; Lovegrove, F.T.

    1984-10-01

    The optimal conditions for labelling infective larvae of Strongyloides ratti with /sup 67/Ga citrate were determined. Radiolabelled larvae were injected s.c. into normal and previously infected rats. The distribution of radioactivity in these animals was compared with that in rats infected subcutaneously with a similar dose of free /sup 67/Ga by using a gamma camera linked to a computer system. Whereas free /sup 67/Ga was distributed throughout the body and excreted via the hepatobiliary system, the bulk of radioactivity in rats injected with radiolabelled larvae remained at the injection sites. Direct microscopical examination of these sites, however, revealed only minimal numbers of worms. When rats were infected percutaneously with radiolabelled larvae, it was found that most radioactivity remained at the surface, despite penetration of worms. When infective larvae were exposed to CO/sub 2/ in vitro and examined carefully by light microscopy, loss of an outer coat was observed. It was concluded that infective larvae lose an outer coat on skin penetration.

  2. Dermal miconazole nitrate nanocrystals - formulation development, increased antifungal efficacy & skin penetration.

    Science.gov (United States)

    Pyo, Sung Min; Hespeler, David; Keck, Cornelia M; Müller, Rainer H

    2017-10-05

    Miconazole nitrate nanosuspension was developed to increase its antifungal activity and dermal penetration. In addition, the nanosuspension was combined with the synergistic additive chlorhexidine digluconate. The production was performed by wet bead milling and both production and formulation parameters were optimized. A stabilizer screening revealed poloxamer 407 and Tween 80 both at 0.15% as the most effective stabilizers for miconazole nanosuspensions at 1.0%. The nanocrystals were incorporated into a hydroxypropyl cellulose gel base. Short-term stability (3months) of the nanocrystal bulk population could be shown at room temperature and fridge. Besides the stable bulk nanocrystals, some longitudinal crystal growth to needle like crystals occurred. The addition of ionic compounds as the chlorhexidine digluconate often destabilizes suspensions. Surprisingly here, the addition minimized the crystal growth. An underlying mechanism is proposed. An inhibition zone assay was performed using Candida albicans (ATCC(®) 10231™). When comparing the nanocrystals in suspension and in gel to μm-sized miconazole nitrate formulations and two market products, the increase in inhibition zone diameter for the nanosuspension formulations was most pronounced in the chlorhexidine digluconate free formulations. These nanocrystal formulations were closely or similarly effective as the microsuspensions and the market products containing the synergistic chlorhexidine digluconate, showing the potential of the nanosuspension formulation. Nanosuspension performance was even further increased when chlorhexidine digluconate was added. Ex-vivo skin penetration studies on porcine ears revealed distinctly less remaining miconazole nitrate on the skin surface for nanocrystals (e.g., 76-86%) compared to market products (e.g. 94%). Also, penetration was increased e.g. in skin depth of 5-10μm from <1.0/1.7% to e.g. 3.3-6.2% for nanocrystals. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Skin penetration of epigallocatechin-3-gallate and quercetin from green tea and Ginkgo biloba extracts vehiculated in cosmetic formulations.

    Science.gov (United States)

    dal Belo, S E; Gaspar, L R; Maia Campos, P M B G; Marty, J-P

    2009-01-01

    Green tea (Camellia sinensis) and Ginkgo biloba extracts in cosmetic formulations have been suggested to protect the skin against UV-induced damage and skin ageing. Thus, it is very important to assess the human skin penetration of their major flavonoids to verify if they penetrate and remain in the skin to exert their proposed effects. The aim of this study was to evaluate the human skin penetration of epigallocatechin-3-gallate (EGCG) and quercetin from green tea and G. biloba extracts vehiculated in cosmetic formulations. This study was conducted with fresh dermatomed human Caucasian skin from abdominal surgery mounted on static Franz diffusion cells. Skin samples were mounted between two diffusion half-cells and 10 mg/cm(2) of formulations supplemented with 6% of green tea or G. biloba extract were applied on the skin surface. The receptor fluid was removed after 6 and 24 h and analyzed by high-performance liquid chromatography for the quantification of the flavonoids. The stratum corneum was removed by tape stripping and immersed in methanol and the epidermis was mechanically separated from the dermis and triturated in methanol to extract EGCG and quercetin. The results showed that the flavonoids under study penetrated into the skin, without reaching the receptor fluid. The majority of EGCG was quantified in the stratum corneum (0.87 microg/cm(2)), which was statistically higher than the EGCG concentrations found in viable epidermis (0.54 microg/cm(2)) and in the dermis (0.38 microg/cm(2)). The majority of quercetin was quantified in the viable epidermis (0.23 microg/cm(2)), which was statistically higher than the EGCG concentration found in the stratum corneum layer (0.17 microg/cm(2)). Finally, it can be concluded that EGCG and quercetin from green tea and G. biloba extracts vehiculated in cosmetic formulations presented good skin penetration and retention, which can favor their skin effects.

  4. Impact of semi-solid formulations on skin penetration of iron oxide nanoparticles

    OpenAIRE

    Musazzi, Umberto M.; Santini, Benedetta; Selmin, Francesca; Marini, Valentina; Corsi, Fabio; Allevi, Raffaele; Ferretti, Anna M.; Prosperi, Davide; Cilurzo, Francesco; Colombo, Miriam; Minghetti, Paola

    2017-01-01

    Background This work aimed to provide useful information on the incidence of the choice of formulation in semi-solid preparations of iron-oxide nanoparticles (IONs). The appropriate analytical methods to assess the IONs physical stability and the effect of the semi-solid preparations on IONs human skin penetration were discussed. The physical stability of IONs (Dh?=?31???4?nm; ??=??65???5?mV) loaded in five semi-solid preparations (0.3% w/v), namely Carbopol gel (CP), hydroxyethyl cellulose g...

  5. Comparative analysis of the effects of CO2 fractional laser and sonophoresis on human skin penetration with 5-aminolevulinic acid.

    Science.gov (United States)

    Choi, J H; Shin, E J; Jeong, K H; Shin, M K

    2017-11-01

    Successful delivery of a photosensitizer into the skin is an important factor for effective photodynamic therapy (PDT). The effective method to increase drug penetration within short incubation time overcoming skin barrier have been investigated. This study was performed to analyze and compare the effectiveness of ablative fractional laser (FXL) pretreatment and/or sonophoresis for enhancing the penetration of 5-aminolevulinic acid (ALA) into human skin in vivo. Twenty-four identical 1 × 1 cm(2) treatment areas were mapped on the backs of ten healthy male subjects. Each area received FXL pretreatment and/or sonophoresis with different energy settings and ALA incubation times. After treatments, porphyrin fluorescence reflecting the ALA penetration were measured. Application of ablative CO2 FXL pretreatment resulted to higher fluorescence intensities than the non-treatment group. Incubation times were positively correlated with the increments of ALA penetration. However, increasing pulse energy or combining with sonophoresis did not show additional positive effects on ALA penetration. Ablative CO2 FXL pretreatment effectively facilitated ALA penetration in human skin in vivo. Ablative CO2 FXL alone without sonophoresis setting pulse energy of 10 and 20 mJ with more than 60 min of ALA incubation time could be an ideal setting for ALA penetration.

  6. Potential use of ascorbic acid-based surfactants as skin penetration enhancers.

    Science.gov (United States)

    Palma, S D; Maletto, B; Lo Nostro, P; Manzo, R H; Pistoresi-Palencia, M C; Allemandi, D A

    2006-08-01

    6-O-Ascorbic acid alkanoates (ASCn) are amphiphilic molecules having physical-chemical properties that depend on the alkyl chain length. The derivatives of low molecular weight (n CMT), ASCn aqueous suspensions turn into either micellar solutions or gel phases, depending on the length of the hydrophobic chain. On cooling, coagels are produced, which possess a lamellar structure that exhibit sharp X-ray diffraction patterns and optical birefringence. The semisolid consistency of such coagels is an interesting property to formulate dermatological pharmaceutical dosage forms able to solubilize and stabilize different drugs. The objective of the present study was the evaluation of the enhancing permeation effect of ASCn with different chain lengths and to correlate permeability changes with histological effects. With this purpose, ASCn coagels containing anthralin (antipsoriasic drug) or fluorescein isothiocyanate (FITC, hydrophobic fluorescent marker) were assayed on rat skin (ex vivo) and mice skin (in vivo), respectively. Also, histological studies were performed aimed at detecting some possible side effects of ASCn. No inflammatory cellular response was observed in the skin when ASCn coagels were applied, suggesting non-irritating properties. Light microscopy indicated slight disruption and fragmentation of stratum corneum. The penetration of ASCn through rat skin epidermis was very fast and quantitatively significant. The permeation of anthralin was significantly increased when the drug was vehiculized in ASCn coagels, compared to other pharmaceutical systems. The results indicated that ASC12 seems to have the highest enhancing effect on FITC permeation. ASC12 appears to be the compound that possesses the highest capacity to enhance the penetration of the drugs. Furthermore, it has the highest permeation of the serie.

  7. In vitro culture of skin-homing T lymphocytes from inflammatory skin diseases

    DEFF Research Database (Denmark)

    Bang, Karen; C Mogensen, Søren; Thestrup-Pedersen, Kristian

    2005-01-01

    was for AD 1.20, MF 0.65 and psoriasis 0.85. Patients with AD treated with cyclosporin-A had almost no growth of CD8+ cells in vitro. Our findings indicate a changed homeostasis among skin-homing lymphocytes for in vitro culture. Our culture system of skin-homing T lymphocytes leads to a prominent cellular......We, in this study, describe how T lymphocytes in a skin biopsy can proliferate in vitro for up to 3 months by using T-cell growth factors - interleukin-2 (IL-2) and IL-4 yielding approximately 100-160 million T lymphocytes within 1 month. We established cell lines from three tuberculin skin tests...

  8. Skin Needling to Enhance Depigmenting Serum Penetration in the Treatment of Melasma

    Science.gov (United States)

    Fabbrocini, G.; De Vita, V.; Fardella, N.; Pastore, F.; Annunziata, M. C.; Mauriello, M. C.; Monfrecola, A.; Cameli, N.

    2011-01-01

    Melasma is a common hypermelanotic disorder affecting the facial area which has a considerable psychological impact on the patient. Managing melasma is a difficult challenge that requires long-term treatment with a number of topical agents, such as rucinol and sophora-alpha. Aims. We aim to compare the combined treatment of skin needling and depigmenting serum with that using depigmenting serum alone in the treatment of melasma, in order to evaluate the use of microneedles as a means to enhance the drug's transdermal penetration. Methods. Twenty patients were treated with combined skin needling and depigmenting serum on one side of the face and with depigmenting serum alone on the other side. The outcome was evaluated periodically for up to two months using the Melasma Area Severity Index score and the Spectrocolorimeter X-Rite 968. Results. The side with combined treatment (skin needling + depigmenting serum) presented a statistically significant reduction in MASI score and luminosity index (L) levels compared to the side treated with depigmenting serum alone, and clinical symptoms were significantly improved. Conclusions. Our study suggests the potential use of combining skin needling with rucinol and sophora-alpha compounds to achieve better results in melasma treatment compared to rucinol and sophora-alpha alone. PMID:22567235

  9. Skin Needling to Enhance Depigmenting Serum Penetration in the Treatment of Melasma

    Directory of Open Access Journals (Sweden)

    G. Fabbrocini

    2011-01-01

    Full Text Available Melasma is a common hypermelanotic disorder affecting the facial area which has a considerable psychological impact on the patient. Managing melasma is a difficult challenge that requires long-term treatment with a number of topical agents, such as rucinol and sophora-alpha. Aims. We aim to compare the combined treatment of skin needling and depigmenting serum with that using depigmenting serum alone in the treatment of melasma, in order to evaluate the use of microneedles as a means to enhance the drug’s transdermal penetration. Methods. Twenty patients were treated with combined skin needling and depigmenting serum on one side of the face and with depigmenting serum alone on the other side. The outcome was evaluated periodically for up to two months using the Melasma Area Severity Index score and the Spectrocolorimeter X-Rite 968. Results. The side with combined treatment (skin needling + depigmenting serum presented a statistically significant reduction in MASI score and luminosity index (L levels compared to the side treated with depigmenting serum alone, and clinical symptoms were significantly improved. Conclusions. Our study suggests the potential use of combining skin needling with rucinol and sophora-alpha compounds to achieve better results in melasma treatment compared to rucinol and sophora-alpha alone.

  10. Skin penetration behavior of lipid-core nanocapsules for simultaneous delivery of resveratrol and curcumin.

    Science.gov (United States)

    Friedrich, Rossana B; Kann, Birthe; Coradini, Karine; Offerhaus, Herman L; Beck, Ruy C R; Windbergs, Maike

    2015-10-12

    Polyphenols, which are secondary plant metabolites, gain increasing research interest due to their therapeutic potential. Among them, resveratrol and curcumin are two agents showing antioxidant, anti-inflammatory, antimicrobial as well as anticarcinogenic effects. In addition to their individual therapeutic effect, increased activity was reported upon co-delivery of the two compounds. However, due to the poor water solubility of resveratrol and curcumin, their clinical application is currently limited. In this context, lipid-core nanocapsules (LNC) composed of an oily core surrounded by a polymeric shell were introduced as drug carrier systems with the potential to overcome this obstacle. Furthermore, the encapsulation of polyphenols into LNC can increase their photostability. As the attributes of the polyphenols make them excellent candidates for skin treatment, the aim of this study was to investigate the effect of co-delivery of resveratrol and curcumin by LNC upon topical application on excised human skin. In contrast to the formulation with one polyphenol, resveratrol penetrated into deeper skin layers when the co-formulation was applied. Based on vibrational spectroscopy analysis, these effects are most likely due to interactions of curcumin and the stratum corneum, facilitating the skin absorption of the co-administered resveratrol. Furthermore, the interaction of LNC with primary human skin cells was analyzed encountering a cellular uptake within 24h potentially leading to intracellular effects of the polyphenols. Thus, the simultaneous delivery of resveratrol and curcumin by LNC provides an intelligent way for immediate and sustained polyphenol delivery for skin disease treatment. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Production of dissolvable microneedles using an atomised spray process: effect of microneedle composition on skin penetration.

    Science.gov (United States)

    McGrath, Marie G; Vucen, Sonja; Vrdoljak, Anto; Kelly, Adam; O'Mahony, Conor; Crean, Abina M; Moore, Anne

    2014-02-01

    Dissolvable microneedles offer an attractive delivery system for transdermal drug and vaccine delivery. They are most commonly formed by filling a microneedle mold with liquid formulation using vacuum or centrifugation to overcome the constraints of surface tension and solution viscosity. Here, we demonstrate a novel microneedle fabrication method employing an atomised spray technique that minimises the effects of the liquid surface tension and viscosity when filling molds. This spray method was successfully used to fabricate dissolvable microneedles (DMN) from a wide range of sugars (trehalose, fructose and raffinose) and polymeric materials (polyvinyl alcohol, polyvinylpyrrolidone, carboxymethylcellulose, hydroxypropylmethylcellulose and sodium alginate). Fabrication by spraying produced microneedles with amorphous content using single sugar compositions. These microneedles displayed sharp tips and had complete fidelity to the master silicon template. Using a method to quantify the consistency of DMN penetration into different skin layers, we demonstrate that the material of construction significantly influenced the extent of skin penetration. We demonstrate that this spraying method can be adapted to produce novel laminate-layered as well as horizontally-layered DMN arrays. To our knowledge, this is the first report documenting the use of an atomising spray, at ambient, mild processing conditions, to create dissolvable microneedle arrays that can possess novel, laminate layering. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Ethosomes - novel vesicular carriers for enhanced delivery: characterization and skin penetration properties.

    Science.gov (United States)

    Touitou, E; Dayan, N; Bergelson, L; Godin, B; Eliaz, M

    2000-04-03

    This work describes a novel carrier for enhanced skin delivery, the ethosomal system, which is composed of phospholipid, ethanol and water. Ethosomal systems were much more efficient at delivering a fluorescent probe to the skin in terms of quantity and depth, than either liposomes or hydroalcoholic solution. The ethosomal system dramatically enhanced the skin permeation of minoxidil in vitro compared with either ethanolic or hydroethanolic solution or phospholipid ethanolic micellar solution of minoxidil. In addition, the transdermal delivery of testosterone from an ethosomal patch was greater both in vitro and in vivo than from commercially available patches. Skin permeation of ethosomal components, ethanol and phospholipid, was demonstrated in diffusion-cell experiments. Ethosomal systems composed of soy phosphatidylcholine 2%, ethanol 30% and water were shown by electron microscopy to contain multilamellar vesicles. 31P-NMR studies confirmed the bilayer configuration of the lipids. Calorimetry and fluorescence measurements suggested that the vesicular bilayers are flexible, having a relatively low T(m) and fluorescence anisotropy compared with liposomes obtained in the absence of ethanol. Dynamic light scattering measurements indicated that ethanol imparted a negative charge to the vesicles. The average vesicle size, as measured by dynamic light scattering, was modulated by altering the ethosome composition. Experiments using fluorescent probes and ultracentrifugation showed that the ethosomes had a high entrapment capacity for molecules of various lyophilicities.

  13. Skin penetration and antioxidant effect of cosmeto-textiles with gallic acid.

    Science.gov (United States)

    Alonso, C; Martí, M; Barba, C; Lis, M; Rubio, L; Coderch, L

    2016-03-01

    In this work, the antioxidant gallic acid (GA) has been encapsulated in microspheres prepared with poly-ε-caprolactone (PCL) and incorporated into polyamide (PA) obtaining the cosmeto-textile. The topical application of the cosmeto-textile provides a reservoir effect in the skin delivery of GA. The close contact of the cosmeto-textile, containing microsphere-encapsulated GA (ME-GA), with the skin and their corresponding occlusion, may be the main reasons that explain the crossing of active principle (GA) through the skin barrier, located in the stratum corneum, and its penetration into the different compartments of the skin, epidermis and dermis. An ex vivo assessment was performed to evaluate the antioxidant effect of the ME-GA on the stratum corneum (SC) using the thiobarbituric acid-reactive species (TBARS) test. The test is based on a non-invasive ex vivo methodology that evaluates lipid peroxides formed in the outermost layers of the SC from human volunteers after UV radiation to determine the effectiveness of an antioxidant. In this case, a ME-GA cosmeto-textile or ME-GA formulation were applied to the skin in vivo and lipid peroxidation (LPO) in the horny layer were determined after UV irradiation. This methodology may be used as a quality control tool to determine ex vivo the percentage of LPO inhibition on human SC for a variety of antioxidants that are topically applied, in this case GA. Results show that LPO formation was inhibited in human SC when GA was applied directly or embedded in the cosmeto-textile, demonstrating the effectiveness of both applications. The percentage of LPO inhibition obtained after both topical applications was approximately 10% for the cosmeto-textile and 41% for the direct application of microspheres containing GA. This methodology could be used to determine the effectiveness of topically applied antioxidants encapsulated in cosmeto-textiles on human SC. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Penetration profile and human cadaver skin distribution of finasteride from vesicular nanocarriers.

    Science.gov (United States)

    Rao, Yuefeng; Zheng, Feiyue; Liang, Xingguang; Wang, Huiyuan; Zhang, Jin; Lu, Xiaoyang

    2015-12-01

    The skin accumulation of therapeutic agents affects the efficiency of topical drug delivery. In this study, in vitro distribution of finasteride of ethosomes and liposomes in human cadaver skin after percutaneous delivery were investigated. Experiments were performed using modified Franz diffusion cells. Finasteride ethosomes, liposomes or hydroethanolic solutions were used as donor medium. Drug distribution at different skin layers and depths were studied by hotplate separation and frozen horizontal slicing technique. The result showed that the accumulation of finasteride in skin ranged from 9.7-24.3 μg/cm(2) at 12 or 24 hours. The ethosomes demonstrated better enhancing ability to deliver finasteride into the dermis layer than liposomes did. The finasteride concentration in the dermis layer from ethosomes was more than sevenfold higher than from liposomes. The finasteride accumulation in ethosomes group showed a distinctive reversed distribution profile. This distinctive reversed distribution profile is meaningful for exerting a favorable pharmacological effect for finasteride. The drug distribution profile in skin layers showed no significant difference between 12 and 24 hours application (p > 0.05). The study demonstrated that finasteride can be accumulated at target site more effectively and maintained at higher level through the application of novel ethosomal carriers.

  15. Study of the vitamins A, E and C esters penetration into the skin by confocal Raman spectroscopy in vivo

    Science.gov (United States)

    Mogilevych, Borys; Isensee, Debora; Rangel, Joao L.; Dal Pizzol, Carine; Martinello, Valeska C. A.; Dieamant, Gustavo C.; Martin, Airton A.

    2015-06-01

    Vitamins A, E and C play important role in skin homeostasis and protection. Hence, they are extensively used in many cosmetic and cosmeceutic products. However, their molecules are unstable, and do not easily penetrate into the skin, which drastically decreases its efficiency in topical formulations. Liposoluble derivative of the vitamin A - retinyl palmitate, vitamin E - tocopheryl acetate, and vitamin C - tetraisopalmitoyl ascorbic acid, are more stable, and are frequently used as an active ingredient in cosmetic products. Moreover, increased hydrophobicity of these molecules could lead to a higher skin penetration. The aim of this work is to track and compare the absorption of the liposoluble derivatives of the vitamins and their encapsulated form, into the healthy human skin in vivo. We used Confocal Raman Spectroscopy (CRS) that is proven to be helpful in label-free non-destructive investigation of the biochemical composition and molecular conformational analysis of the biological samples. The measurements were performed in the volar forearm of the 10 healthy volunteers. Skin was treated with both products, and Raman spectra were obtained after 15 min, 3 hours, and 6 hours after applying the formulation. 3510 Skin Composition Analyzer (River Diagnostics, The Netherlands) with 785 nm laser excitation was used to acquire information in the fingerprint region. Significant difference in permeation of the products was observed. Whereas only free form of retinyl palmitate penetrate the skin within first 15 minutes, all three vitamin derivatives were present under the skin surface in case of nanoparticulated form.

  16. Skin pre-ablation and laser assisted microjet injection for deep tissue penetration.

    Science.gov (United States)

    Jang, Hun-Jae; Yeo, Seonggu; Yoh, Jack J

    2017-04-01

    For conventional needless injection, there still remain many unresolved issues such as the potential for cross-contamination, poor reliability of targeted delivery dose, and significantly painstaking procedures. As an alternative, the use of microjets generated with Er:YAG laser for delivering small doses with controlled penetration depths has been reported. In this study, a new system with two stages is evaluated for effective transdermal drug delivery. First, the skin is pre-ablated to eliminate the hard outer layer and second, laser-driven microjet penetrates the relatively weaker and freshly exposed epidermis. Each stage of operation shares a single Er:YAG laser that is suitable for skin ablation as well as for the generation of a microjet. In this study, pig skin is selected for quantification of the injection depth based on the two-stage procedure, namely pre-ablation and microjet injection. The three types of pre-ablation devised here consists of bulk ablation, fractional ablation, and fractional-rotational ablation. The number of laser pulses are 12, 18, and 24 for each ablation type. For fractional-rotational ablation, the fractional beams are rotated by 11.25° at each pulse. The drug permeation in the skin is evaluated using tissue marking dyes. The depth of penetration is quantified by a cross sectional view of the single spot injections. Multi-spot injections are also carried out to control the dose and spread of the drug. The benefits of a pre-ablation procedure prior to the actual microjet injection to the penetration is verified. The four possible combinations of injection are (a) microjet only; (b) bulk ablation and microjet injection; (c) fractional ablation and microjet injection; and (d) fractional-rotational ablation and microjet injection. Accordingly, the total depth increases with injection time for all cases. In particular, the total depth of penetration attained via fractional pre-ablation increased by 8 ∼ 11% and that of fractional

  17. Microorganism penetration in dentinal tubules of instrumented and retreated root canal walls. In vitro SEM study

    Directory of Open Access Journals (Sweden)

    Saad Al-Nazhan

    2014-11-01

    Full Text Available Objectives This in vitro study aimed to investigate the ability of Candida albicans (C. albicans and Enterococcus faecalis (E. faecalis to penetrate dentinal tubules of instrumented and retreated root canal surface of split human teeth. Materials and Methods Sixty intact extracted human single-rooted teeth were divided into 4 groups, negative control, positive control without canal instrumentation, instrumented, and retreated. Root canals in the instrumented group were enlarged with endodontic instruments, while root canals in the retreated group were enlarged, filled, and then removed the canal filling materials. The teeth were split longitudinally after canal preparation in 3 groups except the negative control group. The teeth were inoculated with both microorganisms separately and in combination. Teeth specimens were examined by scanning electron microscopy (SEM, and the depth of penetration into the dentinal tubules was assessed using the SMILE view software (JEOL Ltd. Results Penetration of C. albicans and E. faecalis into the dentinal tubules was observed in all 3 groups, although penetration was partially restricted by dentin debris of tubules in the instrumented group and remnants of canal filling materials in the retreated group. In all 3 groups, E. faecalis penetrated deeper into the dentinal tubules by way of cell division than C. albicans which built colonies and penetrated by means of hyphae. Conclusions Microorganisms can easily penetrate dentinal tubules of root canals with different appearance based on the microorganism size and status of dentinal tubules.

  18. Nuclear microprobe investigation of the penetration of ultrafine zinc oxide into intact and tape-stripped human skin

    Energy Technology Data Exchange (ETDEWEB)

    Szikszai, Z., E-mail: szikszai@atomki.h [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Kertesz, Zs. [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Bodnar, E. [Department of Dermatology, University of Debrecen, Medical and Health Science Center, Debrecen (Hungary); Major, I. [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Borbiro, I. [Abiol Ltd., Debrecen (Hungary); Kiss, A.Z. [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Hunyadi, J. [Department of Dermatology, University of Debrecen, Medical and Health Science Center, Debrecen (Hungary)

    2010-06-15

    Ultrafine metal oxides, such as titanium dioxide and zinc oxide are widely used in cosmetic and health products like sunscreens. These oxides are potent UV filters and the small particle size makes the product more transparent compared to formulations containing coarser particles. In the present work the penetration of ultrafine zinc oxide into intact and tape-stripped human skin was investigated using nuclear microprobe techniques, such as proton induced X-ray spectroscopy and scanning transmission ion microscopy. Our results indicate that the penetration of ultrafine zinc oxide, in a hydrophobic basis gel with 48 h application time, is limited to the stratum corneum layer of the intact skin. Removing the stratum corneum partially or entirely by tape-stripping did not cause the penetration of the particles into the deeper dermal layers; the zinc particles remained on the surface of the skin.

  19. Novel carbopol-based transfersomal gel of 5-fluorouracil for skin cancer treatment: in vitro characterization and in vivo study.

    Science.gov (United States)

    Khan, Mohammed Ashif; Pandit, Jayamanti; Sultana, Yasmin; Sultana, Sarwat; Ali, Asgar; Aqil, Mohammed; Chauhan, Meenakshi

    2015-01-01

    5-fluorouracil (5-Fu) is an antineoplastic drug, topically used for the treatment of actinic keratosis and nonmelanoma skin cancer. It shows poor percutaneous permeation through the conventionally applicable creams and thus inefficient for the treatment of deep-seated skin cancer. In the present article, transfersomal gel containing 5-Fu was investigated for the treatment of skin cancer. Different formulation of tranfersomes was prepared using Tween-80 and Span-80 as edge activators. The vesicles were characterized for particle size, shape, entrapment efficiency, deformability and in vitro skin permeation. Optimized formulation was incorporated into 1% carbopol 940 gel and evaluated for efficacy in the treatment of skin cancer. 5-Fu-loaded transfersomes (TT-2) has the size of 266.9 ± 2.04 nm with 69.2 ± 0.98% entrapment efficiency and highest deformability index of 27.8 ± 1.08. Formulation TT-2 showed maximum skin deposition (81.3%) and comparable transdermal flux of 21.46 µg/cm(2) h. The TT-2-loaded gel showed better skin penetration and skin deposition of the drug than the marketed formulation. Composition of the transfersomal gel has been proved nonirritant to the skin. We concluded that the developed 5-Fu-loaded transfersomal gel improves the skin absorption of 5-Fu and provide a better treatment for skin cancer.

  20. Analysis of Human and Porcine Skin in vivo/ex vivo for Penetration of Selected Oils by Confocal Raman Microscopy.

    Science.gov (United States)

    Choe, ChunSik; Lademann, Jürgen; Darvin, Maxim E

    2015-01-01

    The subject of oil penetration into the skin is controversially discussed in the scientific literature. Confocal Raman microscopy was used for analyzing oil penetration into the skin. The following methods were applied in the study: methods based on tracking specific peaks (method 1), the nonrestricted multiple least square fit (method 2), analyzing the lipid-to-keratin peak ratio using the perpendicular drop-down cutoff procedure (method 3), and the Gaussian function-based deconvolution procedure (method 4). The results obtained using methods 1, 2 and 4 show that the investigated oils do not penetrate deeper than 11 µm into human and porcine skin. Petrolatum has a prominent swelling effect on the stratum corneum (32% in vivo, 28% ex vivo), while the other oils exhibit no significant swelling effect. By using method 3, the penetration profile of oils, and especially of petrolatum, into the skin was interpreted incorrectly for various reasons that are addressed herein below. Predominantly remaining in the uppermost corneocyte layers of the stratum corneum, topically applied oils do not reach the viable cells of the stratum spinosum. To exclude any possible mistakes when using the lipid-keratin Raman peak (2,820-3,030 cm-1), the penetration analysis should be performed using the Gaussian function-based deconvolution procedure. © 2015 S. Karger AG, Basel.

  1. Vehicle and enhancer effects on human skin penetration of aminophylline from cream formulations: evaluation in vivo.

    Science.gov (United States)

    Wang, Lai-Hao; Wang, Chia-Chen; Kuo, Su-Ching

    2007-01-01

    The effects of four essential oils (rosemary, ylang, lilacin, and peppermint oils), and three plant oils (jojoba oil, corn germ oil, and olive oil) on the permeation of aminophylline were studied using human skin. The permeation effects of these oils were compared with those of three chemical penetration enhancers. Although all oils enhanced the permeation of aminophylline, their effects were less than that of ethanol. Jojoba oil was found to be the most active, causing about a 32% peak height decrease of N-H bending absorbances in comparison with the control, while peppermint, lilacin, rosemary, and ylang oils caused 28%, 24%, 18%, and 12% peak height decreases, respectively. Microemulsions containing 10% jojoba oil and 30% corn germ oil were found to be superior vehicles for the percutaneous absorption of aminophylline. Comparision with results obtained from high-performance liquid chromatography shows good agreement.

  2. Impact of Cosmetic Lotions on Nanoparticle Penetration through ex Vivo C57BL/6 Hairless Mouse and Human Skin: A Comparison Study

    Directory of Open Access Journals (Sweden)

    Samreen Jatana

    2016-02-01

    Full Text Available Understanding the interactions of nanoparticles (NPs with skin is important from a consumer and occupational health and safety perspective, as well as for the design of effective NP-based transdermal therapeutics. Despite intense efforts to elucidate the conditions that permit NP penetration, there remains a lack of translatable results from animal models to human skin. The objectives of this study are to investigate the impact of common skin lotions on NP penetration and to quantify penetration differences of quantum dot (QD NPs between freshly excised human and mouse skin. QDs were mixed in seven different vehicles, including five commercial skin lotions. These were topically applied to skin using two exposure methods; a petri dish protocol and a Franz diffusion cell protocol. QD presence in the skin was quantified using Confocal Laser Scanning Microscopy. Results show that the commercial vehicles can significantly impact QD penetration in both mouse and human skin. Lotions that contain alpha hydroxyl acids (AHA facilitated NP penetration. Lower QD signal was observed in skin studied using a Franz cell. Freshly excised human skin was also studied immediately after the sub-cutaneous fat removal process, then after 24 h rest ex vivo. Resting human skin 24 h prior to QD exposure significantly reduced epidermal presence. This study exemplifies how application vehicles, skin processing and the exposure protocol can affect QD penetration results and the conclusions that maybe drawn between skin models.

  3. Impact of Cosmetic Lotions on Nanoparticle Penetration through ex vivo C57BL/6 Hairless Mouse and Human Skin: A Comparison Study

    Science.gov (United States)

    Jatana, Samreen; Callahan, Linda M.; Pentland, Alice P.; DeLouise, Lisa A.

    2016-01-01

    Understanding the interactions of nanoparticles (NPs) with skin is important from a consumer and occupational health and safety perspective, as well as for the design of effective NP-based transdermal therapeutics. Despite intense efforts to elucidate the conditions that permit NP penetration, there remains a lack of translatable results from animal models to human skin. The objectives of this study are to investigate the impact of common skin lotions on NP penetration and to quantify penetration differences of quantum dot (QD) NPs between freshly excised human and mouse skin. QDs were mixed in 7 different vehicles, including 5 commercial skin lotions. These were topically applied to skin using two exposure methods; a petri dish protocol and a Franz diffusion cell protocol. QD presence in the skin was quantified using Confocal Laser Scanning Microscopy. Results show that the commercial vehicles can significantly impact QD penetration in both mouse and human skin. Lotions that contain alpha hydroxyl acids (AHA) facilitated NP penetration. Lower QD signal was observed in skin studied using a Franz cell. Freshly excised human skin was also studied immediately after the sub-cutaneous fat removal process, then after 24 hours rest ex vivo. Resting human skin 24 hours prior to QD exposure significantly reduced epidermal presence. This study exemplifies how application vehicles, skin processing and the exposure protocol can affect QD penetration results and the conclusions that maybe drawn between skin models. PMID:27453793

  4. Impact of Cosmetic Lotions on Nanoparticle Penetration through ex vivo C57BL/6 Hairless Mouse and Human Skin: A Comparison Study.

    Science.gov (United States)

    Jatana, Samreen; Callahan, Linda M; Pentland, Alice P; DeLouise, Lisa A

    2016-03-01

    Understanding the interactions of nanoparticles (NPs) with skin is important from a consumer and occupational health and safety perspective, as well as for the design of effective NP-based transdermal therapeutics. Despite intense efforts to elucidate the conditions that permit NP penetration, there remains a lack of translatable results from animal models to human skin. The objectives of this study are to investigate the impact of common skin lotions on NP penetration and to quantify penetration differences of quantum dot (QD) NPs between freshly excised human and mouse skin. QDs were mixed in 7 different vehicles, including 5 commercial skin lotions. These were topically applied to skin using two exposure methods; a petri dish protocol and a Franz diffusion cell protocol. QD presence in the skin was quantified using Confocal Laser Scanning Microscopy. Results show that the commercial vehicles can significantly impact QD penetration in both mouse and human skin. Lotions that contain alpha hydroxyl acids (AHA) facilitated NP penetration. Lower QD signal was observed in skin studied using a Franz cell. Freshly excised human skin was also studied immediately after the sub-cutaneous fat removal process, then after 24 hours rest ex vivo. Resting human skin 24 hours prior to QD exposure significantly reduced epidermal presence. This study exemplifies how application vehicles, skin processing and the exposure protocol can affect QD penetration results and the conclusions that maybe drawn between skin models.

  5. Photoacoustic study of the penetration kinetics of nimesulid into human skin

    Energy Technology Data Exchange (ETDEWEB)

    Barja, P R; Veloso, D J D V, E-mail: barja@univap.b [Research and Development Institute, UNIVAP, Av. Shishima Hifumi 2911, Sao Jose dos Campos, SP, 12209-010 (Brazil)

    2010-03-01

    The photoacoustic (PA) effect is observed when modulated (or pulsed) light is absorbed by a sample inside a closed chamber and converted in heat, generating acoustic waves; PA measurements have been employed to evaluate transdermal penetration of topically applied drugs. Phonophoresis is the utilization of ultrasonic (US) energy to enhance absorption of drugs across the epidermal barrier, and its usefulness has been shown by PA measurements. The aim of the present work was to determine the characteristic absorption times of the anti-inflammatory Nimesulid (gel) in human skin, with and without help of therapeutic phonophoresis. After local cleaning, measurements were performed in the forearm of each volunteer before Nimesulid application and for different times after application through massage with the US equipment head; the protocol was repeated for the opposite forearm, but without US emission. Curves of the PA signal level as a function of time were adjusted by a Boltzmann equation, leading to the determination of the characteristic absorption time (about 12 minutes). No significant gain was observed in Nimesulid absorption with the utilization of US radiation, indicating that topic application of Nimesulid does not require the use of phonophoresis, due to the natural fast penetration of the Nimesulid gel.

  6. Investigation of effects of terpene skin penetration enhancers on stability and biological activity of lysozyme.

    Science.gov (United States)

    Varman, Rahul M; Singh, Somnath

    2012-12-01

    The transport of proteins through skin can be facilitated potentially by using terpenes as chemical enhancers. However, we do not know about the effects of these enhancers on the stability and biological activity of proteins which is crucial for the development of safe and efficient formulations. Therefore, this project investigated the effects of terpene-based skin penetration enhancers which are reported as nontoxic to the skin (e.g., limonene, p-cymene, geraniol, farnesol, eugenol, menthol, terpineol, carveol, carvone, fenchone, and verbenone), on the conformational stability and biological activity of a model protein lysozyme. Terpene (5% v/v) was added to lysozyme solution and kept for 24 h (the time normally a transdermal patch remains) for investigating conformational stability profiles and biological activity. Fourier transform infrared spectrophotometer was used to analyze different secondary structures, e.g., α-helix, β-sheet, β-turn, and random coil. Conformational changes were also monitored by differential scanning calorimeter by determining midpoint transition temperature (Tm) and calorimetric enthalpy (ΔH). Biological activity of lysozyme was determined by measuring decrease in A (450) when it was added to a suspension of Micrococcus lysodeikticus. The results of this study indicate that terpenes 9, 10, and 11 (carvone, L-fenchone, and L-verbenone) decreased conformational stability and biological activity of lysozyme significantly (p terpenes used in this study. It is concluded that smaller terpenes containing ketones with low lipophilicity (log K (ow) ∼2.00) would be optimal for preserving conformational stability and biological activity of lysozyme in a transdermal formulation containing terpene as permeation enhancer.

  7. Dermal uptake and percutaneous penetration of ten flame retardants in a human skin ex vivo model

    DEFF Research Database (Denmark)

    Frederiksen, Marie; Vorkamp, Katrin; Jensen, Niels Martin

    2016-01-01

    The dermal uptake and percutaneous penetration of ten organic flame retardants was measured using an ex vivo human skin model. The studied compounds were DBDPE, BTBPE, TBP-DBPE, EH-TBB, BEH-TEBP, α, β and γ-HBCDD as well as syn- and anti-DDC-CO. Little or none of the applied flame retardants...

  8. Simultaneous determination of active component and vehicle penetration from F-DPPC liposomes into porcine skin layers.

    Science.gov (United States)

    Mahrhauser, Denise-Silvia; Reznicek, Gottfried; Gehrig, Sebastian; Geyer, Antonia; Ogris, Manfred; Kieweler, Ruth; Valenta, Claudia

    2015-11-01

    Liposomes have been used as innovative delivery vehicles on skin for a number of years due to their positive influence on skin penetration. However, until now it is not entirely clear how and by which mechanism enhancement is achieved. In the present study, the skin permeation of a model substance incorporated into liposomes and a control formulation was compared to study the influence of the vehicle in Franz-type diffusion cell experiments. Furthermore, the penetration depths of both components were studied by simultaneous determination of the active substance and the vehicle component during tape stripping studies and horizontal sectioning. For these purposes we prepared liposomes with 1-palmitoyl-2-(16-fluoropalmitoyl)-sn-glycero-3-phosphocholine (F-DPPC), the monofluorinated analogue of dipalmitoylphosphaditylcholine (DPPC) loaded with sodium fluorescein (SoFl). A sodium-fluorescein solution was used as control formulation. While the semi-solid F-DPPC liposomes and the SoFl-solution performed equally well with similar permeation profiles during skin diffusion experiments, superior penetrated amounts of SoFl into the stratum corneum (SC) from F-DPPC liposomes compared to the SoFl-solution were observed possibly due to a "push" exerted by the vehicle F-DPPC. We also showed that SoFl penetrated through SC into the viable epidermis. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Effect of rubbing on the in vitro skin permeation of diclofenac-diethylamine 1.16% gel

    Directory of Open Access Journals (Sweden)

    Hasler-Nguyen Nathalie

    2012-06-01

    Full Text Available Abstract Background Rubbing a topical NSAID (non steroidal anti-inflammatory drug on the skin may increase local drug permeation, affecting its distribution to the site of pain and inflammation. The present study evaluates this hypothesis, by assessing in vitro the effect on skin permeation of applying diclofenac-dieythylamine 1.16% gel with or without rubbing. Methods A single dose of 5 mg/cm2 diclofenac-diethylamine 1.16% gel was applied on excised human skin mounted in Franz-type diffusion cells without or with rubbing for 45 s. Drug penetration into the skin layers was determined after 1 h using the tape stripping technique. In vitro cutaneous permeation into the receptor fluid of the diffusion chamber was measured up to 24 h. Skin electrical resistance was also recorded. Results Application of diclofenac-diethylamine 1.16% gel with rubbing resulted to a 5-fold higher flux of diclofenac through the skin than when applied without rubbing at 8 h (P = 0.04. Skin rubbing for 45 s decreased by 2-fold skin electrical resistance when compared to the standard application. Application of diclofenac-diethylamine 1.16% gel with rubbing tended to result in higher accumulation in the stripped skin vs. the superficial skin layers when applied without rubbing (P = 0.2. Conclusion These results suggest that rubbing may alter the superficial skin layer resulting in a transient faster initial diffusion of topically applied diclofenac through the stratum corneum into the deeper skin layer of the dermis to the tissue target.

  10. Nuclear microprobe investigation of the penetration of ultrafine zinc oxide into human skin affected by atopic dermatitis

    Energy Technology Data Exchange (ETDEWEB)

    Szikszai, Z., E-mail: szikszai@atomki.hu [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Kertesz, Zs. [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Bodnar, E. [Department of Dermatology, University of Debrecen, Medical and Health Science Center, Debrecen (Hungary); Borbiro, I. [Abiol Ltd., Debrecen (Hungary); Angyal, A.; Csedreki, L.; Furu, E.; Szoboszlai, Z.; Kiss, A.Z. [Institute of Nuclear Research of the Hungarian Academy of Sciences, Debrecen (Hungary); Hunyadi, J. [Department of Dermatology, University of Debrecen, Medical and Health Science Center, Debrecen (Hungary)

    2011-10-15

    Skin penetration is one of the potential routes for nanoparticles to gain access into the human body. Ultrafine metal oxides, such as titanium dioxide and zinc oxide are widely used in cosmetic and health products like sunscreens. These oxides are potent UV filters and the particle size smaller than 200 nm makes the product more transparent compared to formulations containing coarser particles. The present study continues the work carried out in the frame of the NANODERM: 'Quality of skin as a barrier to ultrafine particles' European project and complements our previous investigations on human skin with compromised barrier function. Atopic dermatitis (a type of eczema) is an inflammatory, chronically relapsing, non-contagious skin disease. It is very common in children but may occur at any age. The exact cause of atopic dermatitis is unknown, but is likely due to a combination of impaired barrier function together with a malfunction in the body's immune system. In this study, skin samples were obtained from two patients suffering from atopic dermatitis. Our results indicate that the ultrafine zinc oxide particles, in a hydrophobic basis gel with an application time of 2 days or 2 weeks, have penetrated deeply into the stratum corneum in these patients. On the other hand, penetration into the stratum spinosum was not observed even in the case of the longer application time.

  11. Nuclear microprobe investigation of the penetration of ultrafine zinc oxide into human skin affected by atopic dermatitis

    Science.gov (United States)

    Szikszai, Z.; Kertész, Zs.; Bodnár, E.; Borbíró, I.; Angyal, A.; Csedreki, L.; Furu, E.; Szoboszlai, Z.; Kiss, Á. Z.; Hunyadi, J.

    2011-10-01

    Skin penetration is one of the potential routes for nanoparticles to gain access into the human body. Ultrafine metal oxides, such as titanium dioxide and zinc oxide are widely used in cosmetic and health products like sunscreens. These oxides are potent UV filters and the particle size smaller than 200 nm makes the product more transparent compared to formulations containing coarser particles. The present study continues the work carried out in the frame of the NANODERM: “Quality of skin as a barrier to ultrafine particles” European project and complements our previous investigations on human skin with compromised barrier function. Atopic dermatitis (a type of eczema) is an inflammatory, chronically relapsing, non-contagious skin disease. It is very common in children but may occur at any age. The exact cause of atopic dermatitis is unknown, but is likely due to a combination of impaired barrier function together with a malfunction in the body's immune system. In this study, skin samples were obtained from two patients suffering from atopic dermatitis. Our results indicate that the ultrafine zinc oxide particles, in a hydrophobic basis gel with an application time of 2 days or 2 weeks, have penetrated deeply into the stratum corneum in these patients. On the other hand, penetration into the stratum spinosum was not observed even in the case of the longer application time.

  12. Immunoarchitectural characterization of a human skin model reconstructed in vitro

    Directory of Open Access Journals (Sweden)

    Luís Ricardo Martinhão Souto

    Full Text Available CONTEXT AND OBJECTIVE: Over the last few years, different models for human skin equivalent reconstructed in vitro (HSERIV have been reported for clinical usage and applications in research for the pharmaceutical industry. Before release for routine use as human skin replacements, HSERIV models need to be tested regarding their similarity with in vivo skin, using morphological (architectural and immunohistochemical (functional analyses. A model for HSERIV has been developed in our hospital, and our aim here was to further characterize its immunoarchitectural features by comparing them with human skin, before it can be tested for clinical use, e.g. for severe burns or wounds, whenever ancillary methods are not indicated. DESIGN AND SETTING: Experimental laboratory study, in the Skin Cell Culture Laboratory, School of Medical Sciences, Universidade Estadual de Campinas. METHODS: Histological sections were stained with hematoxylin-eosin, Masson's trichrome for collagen fibers, periodic acid-Schiff reagent for basement membrane and glycogen, Weigert-Van Gieson for elastic fibers and Fontana-Masson for melanocytes. Immunohistochemistry was used to localize cytokeratins (broad spectrum of molecular weight, AE1/AE3, high molecular weight cytokeratins (34βE12, low molecular weight cytokeratins (35βH11, cytokeratins 7 and 20, vimentin, S-100 protein (for melanocytic and dendritic cells, CD68 (KP1, histiocytes and CD34 (QBend, endothelium. RESULTS: Histology revealed satisfactory similarity between HSERIV and in vivo skin. Immunohistochemical analysis on HSERIV demonstrated that the marker pattern was similar to what is generally present in human skin in vivo. CONCLUSION: HSERIV is morphologically and functionally compatible with human skin observed in vivo.

  13. An in vitro comparison of penetration depth of two root canal sealers: An SEM study

    OpenAIRE

    Chandra Vijay Singh; S Anitha Rao; Chandrashekar, V

    2012-01-01

    Aim: The aim of this study was to examine in vitro penetration depth of two resin-based sealers (AH plus and Resino Seal) and Zinc Oxide Eugenol sealer into the dentinal tubules after removing smear layer by passive ultrasonic irrigation. Materials and Methods: Thirty freshly extracted maxillary central incisors were used. The teeth were decoronated, working length established and prepared upto size 40 file. Each root was subjected to passive ultrasonic irrigation with 2.5% sodium hypoch...

  14. Extrapolation of systemic bioavailability assessing skin absorption and epidermal and hepatic metabolism of aromatic amine hair dyes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Manwaring, John, E-mail: manwaring.jd@pg.com [Procter & Gamble Inc., Mason Business Center, Mason, OH 45040 (United States); Rothe, Helga [Procter & Gamble Service GmbH, Sulzbacher Str. 40, 65823 Schwalbach am Taunus (Germany); Obringer, Cindy; Foltz, David J.; Baker, Timothy R.; Troutman, John A. [Procter & Gamble Inc., Mason Business Center, Mason, OH 45040 (United States); Hewitt, Nicola J. [SWS, Erzhausen (Germany); Goebel, Carsten [Procter & Gamble Service GmbH, Sulzbacher Str. 40, 65823 Schwalbach am Taunus (Germany)

    2015-09-01

    Approaches to assess the role of absorption, metabolism and excretion of cosmetic ingredients that are based on the integration of different in vitro data are important for their safety assessment, specifically as it offers an opportunity to refine that safety assessment. In order to estimate systemic exposure (AUC) to aromatic amine hair dyes following typical product application conditions, skin penetration and epidermal and systemic metabolic conversion of the parent compound was assessed in human skin explants and human keratinocyte (HaCaT) and hepatocyte cultures. To estimate the amount of the aromatic amine that can reach the general circulation unchanged after passage through the skin the following toxicokinetically relevant parameters were applied: a) Michaelis–Menten kinetics to quantify the epidermal metabolism; b) the estimated keratinocyte cell abundance in the viable epidermis; c) the skin penetration rate; d) the calculated Mean Residence Time in the viable epidermis; e) the viable epidermis thickness and f) the skin permeability coefficient. In a next step, in vitro hepatocyte K{sub m} and V{sub max} values and whole liver mass and cell abundance were used to calculate the scaled intrinsic clearance, which was combined with liver blood flow and fraction of compound unbound in the blood to give hepatic clearance. The systemic exposure in the general circulation (AUC) was extrapolated using internal dose and hepatic clearance, and C{sub max} was extrapolated (conservative overestimation) using internal dose and volume of distribution, indicating that appropriate toxicokinetic information can be generated based solely on in vitro data. For the hair dye, p-phenylenediamine, these data were found to be in the same order of magnitude as those published for human volunteers. - Highlights: • An entirely in silico/in vitro approach to predict in vivo exposure to dermally applied hair dyes • Skin penetration and epidermal conversion assessed in human

  15. Influence of the skin barrier on the penetration of topically-applied dexamethasone probed by soft X-ray spectromicroscopy.

    Science.gov (United States)

    Yamamoto, K; Klossek, A; Flesch, R; Rancan, F; Weigand, M; Bykova, I; Bechtel, M; Ahlberg, S; Vogt, A; Blume-Peytavi, U; Schrade, P; Bachmann, S; Hedtrich, S; Schäfer-Korting, M; Rühl, E

    2017-09-01

    The penetration of dexamethasone into human skin ex vivo is reported. X-ray microscopy is used for label-free probing of the drug and quantification of the local drug concentration with a spatial resolution reaching 70±5nm. This is accomplished by selective probing the dexamethasone by X-ray absorption. Varying the penetration time between 10min and 1000min provides detailed information on the penetration process. In addition, the stratum corneum has been damaged by tape-stripping in order to determine the importance of this barrier regarding temporally resolved drug penetration profiles. Dexamethasone concentrations distinctly vary, especially close to the border of the stratum corneum and the viable epidermis, where a local minimum in drug concentration is observed. Furthermore, near the basal membrane the drug concentration strongly drops. High spatial resolution studies along with a de-convolution procedure reveal the spatial distribution of dexamethasone in the interspaces between the corneocytes consisting of stratum corneum lipids. These results on local drug concentrations are interpreted in terms of barriers affecting the drug penetration in human skin. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Sponge like microparticles for drug delivery and cosmeto-textile use: Formulation and human skin penetration.

    Science.gov (United States)

    Zafar, Nadiah; Robin, Sophie; Viennet, Céline; Humbert, Philippe; Valour, Jean Pierre; Agusti, Geraldine; Fessi, Hatem; Elaissari, Abdelhamid

    2017-10-30

    This unique work is targeted to achieve three main goals: i) to enhance the aqueous solubility of three specifically selected hydrophobic active agents, ii) to prepare such polymeric biodegradable microparticles which can encapsulate actives-cyclodextrin complexes and iii) to functionalize a polyamide base textile with active loaded microparticles and active-cyclodextrin loaded microparticles. To achieve this objective, biodegradable cationic microparticles were prepared via double emulsion solvent evaporation process and were loaded with hydroxypropyl-beta-cyclodextrin based complexes of Indomethacin, α-tocopheroland Lauryl Isoquinolinium Bromide during the formulation process. Inclusion complex based particles were evaluated for their morphology, size distribution, zeta potential, skin penetration aptitude and adsorption onto a selected textile. It was observed that active-cyclodextrin complex encapsulation do not affect the morphology, size and zeta potential of the microparticles as well as adsorption of the microparticles onto textile remains unaltered. However such active-cyclodextrin complex encapsulated particles provided the enhancement in the aqueous solubility of hydrophobic agents and also provided prolonged release formulations. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Lecithin-based microemulsions for targeted delivery of ceramide AP into the stratum corneum: formulation, characterizations, and in vitro release and penetration studies.

    Science.gov (United States)

    Sahle, Fitsum F; Metz, Hendrik; Wohlrab, Johannes; Neubert, Reinhard H H

    2013-02-01

    To improve the solubility and penetration of Ceramide AP (CER [AP]) into the stratum corneum that potentially restores the barrier function of aged and affected skin. CER [AP] microemulsions (MEs) were formulated using lecithin, Miglyol® 812 (miglyol) and water-1,2 pentandiol (PeG) mixture as amphiphilic, oily and hydrophilic components, respectively. The nanostructure of the MEs was revealed using electrical conductivity, differential scanning calorimeter (DSC) and electron paramagnetic resonance (EPR) techniques. Photon correlation spectroscopy (PCS) was used to measure the sizes and shape of ME droplets. The release and penetration of the CER into the stratum corneum was investigated in vitro using a multi-layer membrane model. The MEs exhibited excellent thermodynamic stability (>2 years) and loading capacity (0.5% CER [AP]). The pseudo-ternary phase diagrams of the MEs were obtained and PCS results showed that the droplets are spherical in shape and bigger in size. In vitro investigations showed that the MEs exhibited excellent rate and extent of release and penetration. Stable lecithin-based CER [AP] MEs that significantly enhance the solubility and penetration of CER [AP] into the stratum corneum were developed. The MEs also have better properties than the previously reported polyglycerol fatty acid surfactant-based CER [AP] MEs.

  18. Release of arachidonic and linoleic acid metabolites in skin organ cultures as characteristics of in vitro skin irritancy

    NARCIS (Netherlands)

    Sandt, J.J.M. van de; Maas, W.J.M.; Doornink, P.C.; Rutten, A.A.J.J.L.

    1995-01-01

    In vitro techniques make a major contribution to the development of alternatives to the in vivo 'Draize' skin irritation test, and the development of sensitive and generally applicable in vitro endpoints of cutaneous toxicity is an area of intensive research. To investigate in vitro characteristics

  19. Relative Penetration of Zinc Oxide and Zinc Ions into Human Skin after Application of Different Zinc Oxide Formulations.

    Science.gov (United States)

    Holmes, Amy M; Song, Zhen; Moghimi, Hamid R; Roberts, Michael S

    2016-02-23

    Zinc oxide (ZnO) is frequently used in commercial sunscreen formulations to deliver their broad range of UV protection properties. Concern has been raised about the extent to which these ZnO particles (both micronized and nanoparticulate) penetrate the skin and their resultant toxicity. This work has explored the human epidermal skin penetration of zinc oxide and its labile zinc ion dissolution product that may potentially be formed after application of ZnO nanoparticles to human epidermis. Three ZnO nanoparticle formulations were used: a suspension in the oil, capric caprylic triglycerides (CCT), the base formulation commonly used in commercially available sunscreen products; an aqueous ZnO suspension at pH 6, similar to the natural skin surface pH; and an aqueous ZnO suspension at pH 9, a pH at which ZnO is stable and there is minimal pH-induced impairment of epidermal integrity. In each case, the ZnO in the formulations did not penetrate into the intact viable epidermis for any of the formulations but was associated with an enhanced increase in zinc ion fluorescence signal in both the stratum corneum and the viable epidermis. The highest labile zinc fluorescence was found for the ZnO suspension at pH 6. It is concluded that, while topically applied ZnO does not penetrate into the viable epidermis, these applications are associated with hydrolysis of ZnO on the skin surface, leading to an increase in zinc ion levels in the stratum corneum, thence in the viable epidermis and subsequently in the systemic circulation and the urine.

  20. Effect of Penetration Enhancer Containing Vesicles on the Percutaneous Delivery of Quercetin through New Born Pig Skin

    Directory of Open Access Journals (Sweden)

    Maria Manconi

    2011-08-01

    Full Text Available Quercetin (3,3′,4′,5,7-pentahydroxyflavone exerts multiple pharmacological effects: anti-oxidant activity, induction of apoptosis, modulation of cell cycle, anti-mutagenesis, and anti-inflammatory effect. In topical formulations quercetin inhibits oxidative skin damage and the inflammatory processes induced by solar UV radiation. In this work, quercetin (2 mg/mL was loaded in vesicular Penetration Enhancer containing Vesicles (PEVs, prepared using a mixture of lipids (Phospholipon® 50, P50 and one of four selected hydrophilic penetration enhancers: Transcutol® P, propylene glycol, polyethylene glycol 400, and Labrasol® at the same concentration (40% of water phase. Photon Correlation Spectroscopy results showed a mean diameter of drug loaded vesicles in the range 80–220 nm. All formulations showed a negative surface charge and incorporation efficiency in the range 48–75%. Transmission Electron Microscopy confirmed that size and morphology varied as a function of the used penetration enhancer. The influence of PEVs on ex vivo quercetin (transdermal delivery was evaluated using Franz-type diffusion cells, new born pig skin and Confocal Laser Scanning Microscopy. Results showed that drug delivery is affected by the penetration enhancer used in the PEVs' formulation.

  1. Development and validation of an alternative disturbed skin model by mechanical abrasion to study drug penetration

    Science.gov (United States)

    Schlupp, P.; Weber, M.; Schmidts, T.; Geiger, K.; Runkel, F.

    2014-01-01

    Pharmaceuticals and cosmetics for dermal application are usually tested on healthy skin, although the primary permeation barrier, the stratum corneum, is often impaired by skin diseases or small skin lesions, especially on the hands. These skin conditions can considerably influence the permeation of chemicals and drugs. Furthermore, risk assessment for example of nanoparticles should be performed under various skin conditions to reflect the true circumstances. Therefore, an alternative and reproducible method for a high throughput of skin samples with impaired skin barrier was developed and verified by skin permeation studies (25 h) of caffeine, sorbic acid and testosterone compared to healthy (untreated) and tape-stripped skin. Skin barrier disruption was controlled by TEWL measurement. Skin permeation of the three substances was increased in tape-stripped and abraded skin compared to untreated skin due to the reduced barrier integrity. Enhancement of drug uptake was highest for the most hydrophilic substance, caffeine, followed by sorbic acid and lipophilic testosterone. No significant difference in drug uptake studies was observed between the new abrasion method with an aluminum-coated sponge and the tape-stripping method. The obtained results demonstrate that this abrasion method is an alternative way to achieve a disturbed skin barrier for drug and chemical uptake studies. PMID:25756004

  2. In vitro study of the penetration of Streptococcus sanguis and Prevotella intermedia strains into human dentinal tubules.

    Science.gov (United States)

    Berkiten, M; Okar, I; Berkiten, R

    2000-04-01

    The persistent presence of bacteria in the root canal system often leads to the failure of treatment. The purpose of this study was to determine in vitro penetration of Streptococcus sanguis and Prevotella intermedia into dentinal tubules. Samples obtained from human teeth were inoculated with a strain of S. sanguis (NCTC 7853) and P. intermedia (NCTC 93336) for 20 days. Bacterial penetration into tubules was investigated at scanning electron microscopy and light microscopic level. The results showed that S. sanguis could penetrate into dentinal tubules 382.3 microns, whereas P. intermedia could penetrate 25.9 microns. It was observed that P. intermedia had not penetrated into all dentinal tubules. If penetration occurred the depth was quite limited.

  3. In vitro hepatic and skin metabolism of capsaicin.

    Science.gov (United States)

    Chanda, Sanjay; Bashir, Mohammad; Babbar, Sunita; Koganti, Aruna; Bley, Keith

    2008-04-01

    On the basis of the ability of capsaicin to activate the transient receptor potential vanilloid 1 receptor (TRPV1) expressed in nociceptive sensory neurons, topical and injectable high-concentration formulations are being developed as potential treatments for various pain syndromes. As much of the published literature on capsaicin is based on pepper extracts, which are typically a mixture of capsaicin and other capsaicinoids (including norhydrocapsaicin, dihydrocapsaicin, homocapsaicin and homodihydrocapsaicin), the purpose of this investigation was to study the in vitro metabolism of pure capsaicin. The metabolism of capsaicin was similar in human, rat, and dog microsomes and S9 fractions. In these assays, three major metabolites were detected and identified as 16-hydroxycapsaicin, 17-hydroxycapsaicin, and 16,17-dehydrocapsaicin. In addition to these three metabolites, rat microsomes and S9 fractions also produced vanillylamine and vanillin. Biotransformation of capsaicin was slow in human skin in vitro, with the majority of the applied capsaicin remaining unchanged and a small fraction being metabolized to vanillylamine and vanillic acid. These data suggest that the metabolism of capsaicin by cytochrome P450 enzymes in skin is minimal, relative to hepatic metabolism.

  4. An insight into the skin penetration enhancement mechanism of N-methylpyrrolidone.

    Science.gov (United States)

    Cilurzo, Francesco; Vistoli, Giulio; Selmin, Francesca; Gennari, Chiara G M; Musazzi, Umberto M; Franzé, Silvia; Lo Monte, Matteo; Minghetti, Paola

    2014-03-03

    This work aims to elucidate the mechanism by which N-methylpyrrolidone (NMP) enhances the skin permeation of a compound by combining experimental data with molecular dynamic (MD) simulations. The addition of 10% NMP significantly increased the propranolol (PR) permeation through the human epidermis (∼ 15 μg/cm(2) vs ∼ 30 μg/cm(2)) while resulting inefficacious on hydrocortisone (HC) diffusion. No significant alterations in the stratum corneum structure were found after the in vitro treatment of human epidermis with NMP dispersed in mineral oil or water by attenuated total reflectance Fourier transform infrared (ATR-FTIR) analyses. MD simulations revealed the formation of a complex by H-bonds and the π-π stacking interactions between the NMP's amido group and the drug's aromatic systems. The size of the depicted NMP/PR clusters was in line with the hydrodynamic radius derived by dynamic light scattering analyses (∼ 2.00 nm). Conversely, no interaction, and consequently cluster formation, between NMP and HC occurred. These results suggest that NMP is effective in enhancing the drug permeation through human epidermis by a cotransport mechanism when NMP/drug interaction occurs.

  5. Determination of rutin and narcissin in marigold extract and topical formulations by liquid chromatography: applicability in skin penetration studies

    Directory of Open Access Journals (Sweden)

    Yris Maria Fonseca

    2010-01-01

    Full Text Available A chromatographic technique for determination of rutin and narcissin in marigold extract and topical formulations was developed and validated. The method shows linearity over the concentration range of 0.2 - 6.0 μg/mL of rutin (r = 0.9986 and 0.8 - 12.0 μg/mL of narcissin (r = 0.9951. The values obtained for precision and accuracy are in agreement with ICH guidelines. Both the formulation excipients and the porcine ear skin samples did not interfere with the flavonoids determination. The recovery of rutin and narcissin in skin samples added with marigold extract was 81.41% and 83.35%, respectively, which demonstrate the applicability of this method to perform skin penetration studies.

  6. Confocal Raman microscopy and multivariate statistical analysis for determination of different penetration abilities of caffeine and propylene glycol applied simultaneously in a mixture on porcine skin ex vivo.

    Science.gov (United States)

    Mujica Ascencio, Saul; Choe, ChunSik; Meinke, Martina C; Müller, Rainer H; Maksimov, George V; Wigger-Alberti, Walter; Lademann, Juergen; Darvin, Maxim E

    2016-07-01

    Propylene glycol is one of the known substances added in cosmetic formulations as a penetration enhancer. Recently, nanocrystals have been employed also to increase the skin penetration of active components. Caffeine is a component with many applications and its penetration into the epidermis is controversially discussed in the literature. In the present study, the penetration ability of two components - caffeine nanocrystals and propylene glycol, applied topically on porcine ear skin in the form of a gel, was investigated ex vivo using two confocal Raman microscopes operated at different excitation wavelengths (785nm and 633nm). Several depth profiles were acquired in the fingerprint region and different spectral ranges, i.e., 526-600cm(-1) and 810-880cm(-1) were chosen for independent analysis of caffeine and propylene glycol penetration into the skin, respectively. Multivariate statistical methods such as principal component analysis (PCA) and linear discriminant analysis (LDA) combined with Student's t-test were employed to calculate the maximum penetration depths of each substance (caffeine and propylene glycol). The results show that propylene glycol penetrates significantly deeper than caffeine (20.7-22.0μm versus 12.3-13.0μm) without any penetration enhancement effect on caffeine. The results confirm that different substances, even if applied onto the skin as a mixture, can penetrate differently. The penetration depths of caffeine and propylene glycol obtained using two different confocal Raman microscopes are comparable showing that both types of microscopes are well suited for such investigations and that multivariate statistical PCA-LDA methods combined with Student's t-test are very useful for analyzing the penetration of different substances into the skin. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Colloidal carriers of isotretinoin for topical acne treatment: skin uptake, ATR-FTIR and in vitro cytotoxicity studies.

    Science.gov (United States)

    Gürbüz, Aslı; Özhan, Gül; Güngör, Sevgi; Erdal, M Sedef

    2015-09-01

    Acne vulgaris is the chronical, multifactorial and complex disease of the pilosebaceous unit in the skin. The main goal of the topical therapy in acne is to target the drug to epidermal and deep dermal regions by minimizing systemic absorption . Isotretinoin, a retinoic acid derivative, is the most effective drug in acne pathogenesis. Because systemic treatment may cause many side effects, topical isotretinoin treatment is an option in the management of acne. However, due to its high lipophilic character, isotretinoin tends to accumulate in the upper stratum corneum, thus its penetration into the lower layers is limited, which restricts the efficiency of topical treatment. Microemulsions are fluid, isotropic, colloidal drug carriers that have been widely studied as drug delivery systems. The percutaneous transport of active agents can be enhanced by microemulsions when compared with their conventional formulations. The purpose of this study was to evaluate microemulsions as alternative topical carriers for isotretinoin with an objective to improve its skin uptake. After in vitro permeation studies, the dermal penetration of isotretinoin from microemulsions was investigated by tape stripping procedure. Confocal laser scanning microscopy provided insight about the localization of the drug in the skin. The interaction between the microemulsion components and stratum corneum lipids is studied by ATR-FTIR spectroscopy. The relative safety of the microemulsions was assessed in mouse embryonic fibroblasts using MTT viability test. The results indicate that microemulsion-based novel colloidal carriers have a potential for enhanced skin delivery and localization of isotretinoin.

  8. Dilution of semi-solid creams: influence of various production parameters on rheological properties and skin penetration.

    Science.gov (United States)

    Nagelreiter, C; Kratochvilova, E; Valenta, C

    2015-01-30

    In order to customise treatment for patients, topical formulations are often diluted with drug-free cream bases to adjust the drug dose and thereby the formulations' activity to the patients' needs. However, the process of dilution influences properties of the formulations. Stability can be reduced as well as the microbial stability and most importantly, efficacy and skin penetration behaviour can be severely and unpredictably changed. The present study investigates the effects of production parameters on creams, namely incorporation of an API (active pharmaceutical ingredients) into an OW cream with prior mixing with propylene glycol or without and subsequent automated or manual dilution of the resulting creams with three different cream bases. Effects were measured by influence on microscopic appearance, measurement of chemical stability, skin penetration and rheological behaviour. suggest strong influence of the cream bases used for dilution of the formulations. Mixture of equal amounts of the employed OW and WO cream proved unfavourable due to inferior penetration behaviour and less appealing microscopic and macroscopic appearance. Prior mixing with PG was of negligible importance for the characteristics of the dilutions, however, the type of API and manner of dilution had an influence on the viscosity of the formulations. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Dermal penetration and resorption of beta-naphthylamine and N-phenyl-beta-naphthylamine from lubricants in an ex vivo human skin model.

    Science.gov (United States)

    Dennerlein, Kathrin; Göen, Thomas; Zobel, Melanie; Boos, Anja M; Drexler, Hans; Kilo, Sonja

    2017-10-01

    Dermal Penetration of aromatic amines (AA's), often suspected or known to be carcinogenic, can play an important role in the overall human exposure. However, information on penetration of certain AA's is poor and inconsistent. Penetration of the former lubricant additive N-phenyl-beta-naphthylamine (PBNA) and its contaminant beta-naphthylamine (BNA) a known carcinogen was investigated and the influence of formulation and co-application characterized. Percutaneous penetration of BNA and PBNA through freshly excised human skin (n = 8; 48 h) was investigated using an ex vivo diffusion cell model. Both AA's were applied in a technical-conform lubricant or dissolved in hexane. The amount of BNA and PBNA applied to skin was 0.52 and 259 μg/0.64 cm(2). The analytical determination of AA's was performed by GC-MS. Both, BNA and PBNA penetrated through human skin (38 vs. 5% of applied dose). In contrast to BNA, the percutaneous penetration of PBNA continued beyond the end of exposure. Co-exposure of both AA's increased the intradermal uptake of BNA and PBNA (p penetration (2.9 and 1.9% of applied dose). The results clearly reveal that dermal penetration of both AA's depends strongly on the mode of application. Co-application and formulation alters the penetration of the AA's. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Fast profiling ecotoxicity and skin permeability of benzophenone ultraviolet filters using biopartitioning micellar chromatography based on penetrable silica spheres.

    Science.gov (United States)

    Yin, Chen-ru; Ma, Li-yun; Huang, Jian-geng; Xu, Li; Shi, Zhi-guo

    2013-12-04

    Penetrable silica possesses hierarchical pores, mesopores and penetrable macropores, offering fast mass transfer, satisfactory mechanical strength as well as low column pressure. In the present study, penetrable octadecyl-bonded silica (ODS) was for the first time used as biopartitioning micellar chromatography (BMC) stationary phase to profile ecotoxicity and skin permeability of benzophenone UV-filters. Mobile phase (MP) pH and concentration of polyoxyethylene(23)lauryl ether in the MP were systematically studied. Quantitative retention-activity relationships (QRARs) model was established to correlate retention factors (k) on BMC with bioconcentration factor (BCF) and transdermal rate (TR) of UV-filters. Coefficient of determination (r(2)) of the QRARs model between log BCF and log k were 0.9398-0.9753, while r(2) between TR and log k were 0.7569-0.8434, which demonstrated satisfactory predictive ability of the methodology. It was a powerful tool for fast screening by combining penetrable ODS with BMC, and avoiding column blockage often occurring in BMC. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. How Confocal Is Confocal Raman Microspectroscopy on the Skin? Impact of Microscope Configuration and Sample Preparation on Penetration Depth Profiles.

    Science.gov (United States)

    Lunter, Dominique Jasmin

    2016-01-01

    The aim of the study was to elucidate the effect of sample preparation and microscope configuration on the results of confocal Raman microspectroscopic evaluation of the penetration of a pharmaceutical active into the skin (depth profiling). Pig ear skin and a hydrophilic formulation containing procaine HCl were used as a model system. The formulation was either left on the skin during the measurement, or was wiped off or washed off prior to the analysis. The microscope configuration was varied with respect to objectives and pinholes used. Sample preparation and microscope configuration had a tremendous effect on the results of depth profiling. Regarding sample preparation, the best results could be observed when the formulation was washed off the skin prior to the analysis. Concerning microscope configuration, the use of a 40 × 0.6 numerical aperture (NA) objective in combination with a 25-µm pinhole or a 100 × 1.25 NA objective in combination with a 50-µm pinhole was found to be advantageous. Complete removal of the sample from the skin before the analysis was found to be crucial. A thorough analysis of the suitability of the chosen microscope configuration should be performed before acquiring concentration depth profiles. © 2016 S. Karger AG, Basel.

  12. Development and validation of an alternative disturbed skin model by mechanical abrasion to study drug penetration

    Directory of Open Access Journals (Sweden)

    P. Schlupp

    2014-01-01

    Skin permeation of the three substances was increased in tape-stripped and abraded skin compared to untreated skin due to the reduced barrier integrity. Enhancement of drug uptake was highest for the most hydrophilic substance, caffeine, followed by sorbic acid and lipophilic testosterone. No significant difference in drug uptake studies was observed between the new abrasion method with an aluminum-coated sponge and the tape-stripping method. The obtained results demonstrate that this abrasion method is an alternative way to achieve a disturbed skin barrier for drug and chemical uptake studies.

  13. Three-Dimensional In Vitro Skin and Skin Cancer Models Based on Human Fibroblast-Derived Matrix.

    Science.gov (United States)

    Berning, Manuel; Prätzel-Wunder, Silke; Bickenbach, Jackie R; Boukamp, Petra

    2015-09-01

    Three-dimensional in vitro skin and skin cancer models help to dissect epidermal-dermal and tumor-stroma interactions. In the model presented here, normal human dermal fibroblasts isolated from adult skin self-assembled into dermal equivalents with their specific fibroblast-derived matrix (fdmDE) over 4 weeks. The fdmDE represented a complex human extracellular matrix that was stabilized by its own heterogeneous collagen fiber meshwork, largely resembling a human dermal in vivo architecture. Complemented with normal human epidermal keratinocytes, the skin equivalent (fdmSE) thereof favored the establishment of a well-stratified and differentiated epidermis and importantly allowed epidermal regeneration in vitro for at least 24 weeks. Moreover, the fdmDE could be used to study the features of cutaneous skin cancer. Complementing fdmDE with HaCaT cells in different stages of malignancy or tumor-derived cutaneous squamous cell carcinoma cell lines, the resulting skin cancer equivalents (fdmSCEs) recapitulated the respective degree of tumorigenicity. In addition, the fdmSCE invasion phenotypes correlated with their individual degree of tissue organization, disturbance in basement membrane organization, and presence of matrix metalloproteinases. Together, fdmDE-based models are well suited for long-term regeneration of normal human epidermis and, as they recapitulate tumor-specific growth, differentiation, and invasion profiles of cutaneous skin cancer cells, also provide an excellent human in vitro skin cancer model.

  14. Organic Pollutant Penetration through Fruit Polyester Skin: A Modified Three-compartment Diffusion Model

    Science.gov (United States)

    Li, Yungui; Li, Qingqing; Chen, Baoliang

    2016-03-01

    The surface of plants is covered by a continuous but heterogeneous cuticular membrane (CM). Serving as the first protective barrier, the uptake and transport behavior of organic pollutants at this interface continue to engage the research efforts of environmental chemist. To date, the contributions of cuticular components as a defense against the organic pollutants penetration remain unresolved. In this study, the unsteady-state penetration characteristics of phenanthrene (PHE) through isolated fruit CM was investigated. PHE penetration was differentiated by three cuticular compartments: epicuticular waxes (EW), cuticle proper (CP) and cuticular layer (CL). The driving force for PHE penetration was ascribed to the sharp concentration gradient built up endogenously by cuticular compartments with different lipophilic affinities. A modified penetration model was established and verified in terms of its general suitability for the hydrophobic chemicals and CMs of various plant species (apple, tomato and potato). The new three-compartment model demonstrates much higher accuracy in characterizing the uptake and transport behavior of semivolatile chemicals with fewer limitations in terms of environmental conditions and complexity (e.g., coexisting contaminants and temperature). This model could contribute to a more comprehensive understanding on the role of polymeric lipids in the organic pollutant sorption and transport into plants.

  15. Association between microbial flora and tissue abnormality around dental implants penetrating the skin in reconstructed oral cancer patients.

    Science.gov (United States)

    Ahmed, Ayman; Chambers, Mark S; Goldschmidt, Millicent C; Habib, Ahmed; Lei, Xiudong; Jacob, Rhonda F

    2012-01-01

    Patients with skin-penetrating implants sometimes report episodic soft tissue reactions. This pilot clinical trial identified the oral microflora of the peri-implant sulcus of cancer patients with jaw reconstruction and compared sites with and without clinical signs of inflammation. Ten patients were selected during routine follow-up of their implant-supported removable prostheses. Eligible patients had at least two intraoral skin-penetrating implants; one showed clinical signs of inflammation with/without symptoms and the other served as a healthy control site. Eight of the 10 patients had undergone osseocutaneous fibula free flap reconstruction and two had received split-thickness skin grafts on the surface of the native mandible. Subjects were assessed on two visits, 30 to 40 days apart. Subgingival microbial samples were obtained and periodontal pocket depths were measured at the test implant abutment. Between visits, patients followed a strict oral hygiene regimen. Radiographic marginal bone loss around implants was measured using cone beam imaging during the second visit. The microflora were identified after isolation and purification of all growing colonies. The number of different microorganisms ranged from 10 to 21 per site. Among all patients, a total of 47 different microorganisms were identified, but none were considered virulent red-complex periodontal pathogens. By the time of the second visit, after adherence to strict mechanical oral hygiene, the average number of microorganisms per site was reduced through elimination of some species. Almost all subjects had identical microbial species in both sites, but the number of visible colonies was lower at the healthy site. Oral peri-implant epidermal proliferation does not appear to be caused by specific microbial pathogens but is likely related to increased microbial load in the implant sulci. Mechanical oral hygiene improves and maintains healthier peri-implant skin tissues.

  16. Topical delivery of 5-aminolevulinic acid-encapsulated ethosomes in a hyperproliferative skin animal model using the CLSM technique to evaluate the penetration behavior.

    Science.gov (United States)

    Fang, Yi-Ping; Huang, Yaw-Bin; Wu, Pao-Chu; Tsai, Yi-Hung

    2009-11-01

    Psoriasis, an inflammatory skin disease, exhibits recurring itching, soreness, and cracked and bleeding skin. Currently, the topical delivery of 5-aminolevulinic acid-photodynamic therapy (ALA-PDT) is an optional treatment for psoriasis which provides long-term therapeutic effects, is non-toxic and enjoys better compliance with patients. However, the precursor of ALA is hydrophilic, and thus its ability to penetrate the skin is limited. Also, little research has provided a platform to investigate the penetration behavior in disordered skin. We employed a highly potent ethosomal carrier (phosphatidylethanolamine; PE) to investigate the penetration behavior of ALA and the recovery of skin in a hyperproliferative murine model. We found that the application of ethosomes produced a significant increase in cumulative amounts of 5-26-fold in normal and hyperproliferative murine skin samples when compared to an ALA aqueous solution; and the ALA aqueous solution appeared less precise in terms of the penetration mode in hyperproliferative murine skin. After the ethosomes had been applied, the protoporphyrin IX (PpIX) intensity increased about 3.64-fold compared with that of the ALA aqueous solution, and the penetration depth reached 30-80 microm. The results demonstrated that the ethosomal carrier significantly improved the delivery of ALA and the formation of PpIX in both normal and hyperproliferative murine skin samples, and the expression level of tumor necrosis factor (TNF)-alpha was reduced after the ALA-ethosomes were applied to treat hyperproliferative murine skin. Furthermore, the results of present study encourage more investigations on the mechanism of the interaction with ethosomes and hyperproliferative murine skin.

  17. Novel insights for permeant lead structures through in vitro skin diffusion assays of Prunus lusitanica L., the Portugal Laurel

    Science.gov (United States)

    Costa, Maria do Céu; Duarte, Patrícia; Neng, Nuno R.; Nogueira, José M. F.; Costa, Filomena; Rosado, Catarina

    2015-01-01

    As a contribution for the generation of libraries in which a natural product (NP) is used as the guiding structure, this work sought to investigate molecular features of triterpenes as deliver leads to cross the stratum corneum at a significant rate. Seeking a bioguided investigation of the dermocosmetic lead-like potential of triterpenes in Prunus lusitanica L., various extracts were obtained by two different methods (Soxhlet extractor and Accelerated Solvent Extraction-ASE) and analyzed by GC-MS and NMR. In vitro assays were conducted to quantify the friedelin 1 and crude plant extract permeation through a membrane of polydimethylsiloxane (PDMS), as well as their skin penetration enhancement capacity using two model molecules, caffeine 19 and ibuprofen 20. Friedelin 1 was identified as the major component (16-77%, GC) with isolated yield of 51% w/w (94%, GC) from Soxhlet residue (1.7% p/p) of the dried aerial parts of the plant harvested when in early flowering stage. Friedelin 1 promoted the penetration of the lipophilic molecule 20, however, it did not influence the permeation of the hydrophilic permeant 20. On the other hand, the crude extract acted as a retardant of the penetration of both substances. Molecular characteristics for the applicability of P. lusitanica L. in the development of dermocosmetics, as well as a new potential use for friedelin 1 in particular, are demonstrated. Probable mechanisms for chemical penetration enhancement using triterpenes as models for transdermal administration are herein discussed.

  18. Pre-treatment with Aloe vera juice does not enhance the in vitro permeation of ketoprofen across skin.

    Science.gov (United States)

    Ballam, L; Heard, C M

    2010-01-01

    The potential of pre-treating skin with Aloe vera juice as a penetration enhancer was evaluated in vitro using ketoprofen as model permeant. To excised porcine skin mounted in Franz diffusion cells was applied either: (1) commercial Aloe vera; (2) commercial Aloe vera followed by massaging; (3) previously boiled commercial Aloe vera; (4) water (negative control); (5) tea tree oil (positive control). After 1 h, the pre-treatment was removed and the skin dosed with a saturated solution of ketoprofen in polyethylene glycol 400; the appearance of drug in the receptor phase was then monitored by HPLC. No statistically significant differences in the transdermal delivery of ketoprofen were observed between water and all the Aloe vera pre-treatments (p > 0.05). The tea tree oil pre-treatment was significantly different to all others (p Aloe vera appears to have no value as a penetration enhancer when used as a pre-treatment, although the data indirectly support the mechanism of action proposed previously, work when used 'within-vehicle'. Handling household products containing Aloe vera appears not to leave the user at elevated risk of subsequent absorption of exogenous chemicals. (c) 2009 S. Karger AG, Basel.

  19. In vitro Percutaneous Absorption of Niacinamide and Phytosterols and in vivo Evaluation of their Effect on Skin Barrier Recovery.

    Science.gov (United States)

    Offerta, Alessia; Bonina, Francesco; Gasparri, Franco; Zanardi, Andrea; Micicche, Lucia; Puglia, Carmelo

    2016-01-01

    In this study, we evaluated different strategies to optimize the percutaneous absorption of niacinamide (NA) and soy phytosterols (FITO) by making use of solid lipid nanoparticles (SLN) and penetration enhancers, such as the hydrogenated lecithin. The evaluation of the skin permeation of NA and FITO has been effected in vitro using excised human skin (i.e., stratum corneum-epidermis or SCE). Furthermore, we evaluated the in vivo effect that NA and FITO has on skin barrier recovery after the topical application; using the extent of methyl nicotinate (MN)-induced erythema in damaged skin as a parameter to determine the rate of stratum corneum recovery. Results pointed out the importance of these strategies as valid tools for NA and FITO topical delivery. In fact, soy lecithin based formulations were able to increase the percutaneous absorption of the two active ingredients, while SLN guaranteed an interesting delayed and sustained release of FITO. In vivo evaluation showed clearly that the formulation containing both the actives (NA and FITO) is able to recover about 95% of skin barrier integrity eight days after tape stripping. This effect is probably due to the "synergistic effect" of NA and FITO.

  20. A cyclohexanecarboxamide derivative with inhibitory effects on Schistosoma mansoni cercarial serine protease and penetration of mice skin by the parasite.

    Science.gov (United States)

    Bahgat, Mahmoud; Aboul-Enein, Mohamed N; El Azzouny, Aida A; Maghraby, Amany; Ruppel, Andreas; Soliman, Wael M

    2009-01-01

    A cyclohexanecarboxamide derivative, N-phenyl-N-[1-(piperidine-1-carbonyl)cyclohexyl] benzamide (MNRC-5), was evaluated for its inhibitory effects on Schistosoma mansoni cercarial serine protease activity and cercarial penetration. MNRC-5 exerted an inhibitory effect on S. mansoni cercarial serine protease at serial concentrations of the specific chromogenic substrate Boc-Val-Leu-Gly-Arg-PNA for such enzyme family and the inhibitory coefficient (Ki) value was deduced. Moreover, topical treatment of mice tails with the most potent inhibitory concentration of MNRC-5 formulated in jojoba oil successfully blocked cercarial penetration as demonstrated by a significant reduction (75%; p jojoba oil base containing no MNRC-5. In addition, the IgM and IgG reactivities to crude S. mansoni cercarial, worm and egg antigens were generally lower in sera from treated infected mice than untreated infected mice. In conclusion, we report on a new serine protease inhibitor capable for blocking penetration of host skin by S. mansoni cercariae as measured by lowering worm burden and decrease in the levels of both IgM and IgG towards different bilharzial antigens upon topical treatment.

  1. Monte Carlo study of skin optical clearing to enhance light penetration in the tissue: implications for photodynamic therapy of acne vulgaris

    Science.gov (United States)

    Bashkatov, Alexey N.; Genina, Elina A.; Tuchin, Valery V.; Altshuler, Gregory B.; Yaroslavsky, Ilya V.

    2008-06-01

    Result of Monte Carlo simulations of skin optical clearing is presented. The model calculations were carried out with the aim of studying of spectral response of skin under immersion liquids action and calculation of enhancement of light penetration depth. In summary, we have shown that: 1) application of glucose, propylene glycol and glycerol produced significant decrease of light scattering in different skin layers; 2) maximal clearing effect will be obtained in case of optical clearing of skin dermis, however, absorbed light fraction in skin dermis changed insignificantly, independently on clearing agent and place it administration; 3) in contrast to it, the light absorbed fraction in skin adipose layer increased significantly in case of optical clearing of skin dermis. It is very important because it can be used for development of optical methods of obesity treatment; 4) optical clearing of superficial skin layers can be used for decreasing of power of light radiation used for treatment of acne vulgaris.

  2. Anti-MMP-2 Activity and Skin-Penetrating Capability of the Chemical Constituents from Rhodiola rosea.

    Science.gov (United States)

    Lee, Tzong-Huei; Hsu, Chieh-Chih; Hsiao, George; Fang, Jia-You; Liu, Wei-Min; Lee, Ching-Kuo

    2016-05-01

    Based on the significant inhibitory activity toward matrix metalloproteinase-2 and collagenase noticed in preliminary studies, crude extracts of Rhodiola rosea were partitioned and chromatographed sequentially to afford three new compounds, 1,2,3,6-tetra-O-galloyl-4-O-p-hydroxybenzoyl-β-D-glucopyranoside (1), (E)-creoside I (2), and (R,Z)-2-methylhept-2-ene-1,6-diol (3), along with twenty-four known compounds (4-27). Their structures were determined by spectroscopic data analyses. All isolated compounds were subjected to bioactivity assays. In these, 1 specifically inhibited matrix metalloproteinase-2 activity with an IC50 value of 16.3 ± 1.6 µM, while its analogue 1,2,3,6-tetra-O-galloyl-β-D-glucopyranonoside (17) inhibited matrix metalloproteinase-2 with an IC50 value of 23.0 ± 4.8 µM. In the collagenase activity assay, the inhibitory effects of 1 and 17 at concentrations of both 20 and 40 µM were more potent than those of the positive control, 1,10-phenanthroline. In order to realize whether 17 could penetrate from the epidermal layer into the basal and dermal layers of the human skin to inhibit the activity of matrix metalloproteinase-2 and collagenase or not, a transdermal penetration test in nude and white mice skins was performed. Penetration percentages of 17 quantified by LC-MS were 27.8 % and 74.8 % in 24 hours, respectively. Georg Thieme Verlag KG Stuttgart · New York.

  3. In vitro evaluation of copaiba oil as a kojic acid skin enhancer

    Directory of Open Access Journals (Sweden)

    Robson Vicente Machado de Oliveira

    2010-06-01

    Full Text Available The capacity of copaíba oil to act as a skin penetration enhancer for the depigmenting agent kojic acid was evaluated using an in vitro diffusion system with static flux and shed rattlesnake skin membrane, Crotalus durissus terrificus, in saline solution at 34±2 ºC as the fluid receptor. The quantities of kojic acid liberated into the fluid receptor were determined by spectrophotometry at 268 nm with intervals of one and a half hours. The membranes, pretreated with copaíba oil at 25% and 50% v/v, gave flux values of 8.0 and 12.7 µg/cm²/h, permeability values of 2.0 and 3.3 cm×10-4/h, and promotion factors of 4.1 and 3.7, respectively. These results indicate that copaíba oil, at the two concentrations studied, has the capacity to promote penetration of kojic acid.A propriedade do óleo de copaíba como agente promotor de penetração cutânea do despigmentante ácido kójico foi avaliada utilizando-se sistema de difusão in vitro com fluxo estático, membrana de pele da serpente cascavel - Crotalus durissus terrificus e solução salina a 34±2 ºC como fluido receptor. As quantidades liberadas do ácido kójico no fluido receptor foram determinadas por espectrofotometria em 268 nm em intervalos de 1:30 h. As membranas pré-tratadas com óleo de copaíba a 25 e 50% v/v apresentaram valores de fluxo de 8,0 e 12,7 µg/cm²/h, permeabilidade de 2,0 e 3,3 cm×10-4/h, e fatores de promoção de 4,1 e 3,7, respectivamente. Os resultados obtidos indicaram que o óleo de copaíba, nas duas concentrações estudadas, apresentou capacidade de promoção da penetração do ácido kójico.

  4. Oligogalacturonides improve tissue organization of in vitro reconstructed skin.

    Science.gov (United States)

    Lebreton-Decoster, C; Rousselle, P; Laperdrix, C; Lubrano, C; Robin, J-R; Coulomb, B

    2011-10-01

    The aim of this study was to analyse the effects of oligogalacturonides obtained from apple pectin enzymatic hydrolysis (mainly composed of galacturonic acid and oligogalacturonides; OGA) on normal human keratinocytes behaviour using different in vitro models. We demonstrate that 0.01% OGA promotes epidermal growth, organization and stratification in an in vitro reconstructed skin. The presence and the in vivo-like location of epidermal differentiation markers (i.e. keratin 10, involucrin, desmoglein 1 and 3, and cathepsin D) confirms the histological analysis, and underlines the cohesion of the treated epidermis. On the opposite, 0.05% OGA delays epidermal growth and disturbs differentiation, showing that the positive effects of OGA are dependent on its concentration. In parallel, using collagen IV and laminin 332 substrates, two relevant components of dermal-epidermal basement membrane, we demonstrate that the presence of 0.01% OGA clearly stimulates keratinocytes spreading out, paralleled by a well-organized microfilament network. Keratinocytes develop more focal adhesions with the substrates, implicating α6β4 on laminin 332. Cellular cohesion is also promoted by 0.01% OGA through the over-expression of integrins α2β1 on collagen IV, and α3β1 on laminin 332 at cell-cell junctions. Thus, by modulating integrins expression and organization, OGA 0.01% should improve cell-cell interactions and therefore dermal-epidermal cohesion. In conclusion, 0.01% OGA stimulates epidermal spreading and promotes keratinocytes attachment to basement membrane components by reorganizing cytoskeleton and modulating integrins recruitment. Furthermore, 0.01% OGA promotes epidermal differentiation and regulates epidermis homeostasis. Considering that OGA has a beneficial effect on parameters playing a key role in ageing, OGA can be presented as a new anti-ageing active ingredient. © 2011 The Authors. ICS © 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  5. Titanium Dioxide Nanoparticle Penetration into the Skin and Effects on HaCaT Cells

    Directory of Open Access Journals (Sweden)

    Matteo Crosera

    2015-08-01

    Full Text Available Titanium dioxide nanoparticles (TiO2NPs suspensions (concentration 1.0 g/L in synthetic sweat solution were applied on Franz cells for 24 h using intact and needle-abraded human skin. Titanium content into skin and receiving phases was determined. Cytotoxicity (MTT, AlamarBlue® and propidium iodide, PI, uptake assays was evaluated on HaCat keratinocytes after 24 h, 48 h, and seven days of exposure. After 24 h of exposure, no titanium was detectable in receiving solutions for both intact and damaged skin. Titanium was found in the epidermal layer after 24 h of exposure (0.47 ± 0.33 μg/cm2 while in the dermal layer, the concentration was below the limit of detection. Damaged skin, in its whole, has shown a similar concentration (0.53 ± 0.26 μg/cm2. Cytotoxicity studies on HaCaT cells demonstrated that TiO2NPs induced cytotoxic effects only at very high concentrations, reducing cell viability after seven days of exposure with EC50s of 8.8 × 10−4 M (MTT assay, 3.8 × 10−5 M (AlamarBlue® assay, and 7.6 × 10−4 M (PI uptake, index of a necrotic cell death. Our study demonstrated that TiO2NPs cannot permeate intact and damaged skin and can be found only in the stratum corneum and epidermis. Moreover, the low cytotoxic effect observed on human HaCaT keratinocytes suggests that these nano-compounds have a potential toxic effect at the skin level only after long-term exposure.

  6. Synergistic efficacy of salicylic acid with a penetration enhancer on human skin monitored by OCT and diffuse reflectance spectroscopy

    Science.gov (United States)

    Zhao, Qingliang; Dai, Cuixia; Fan, Shanhui; Lv, Jing; Nie, Liming

    2016-10-01

    Salicylic acid (SA) has been frequently used as a facial chemical peeling agent (FCPA) in various cosmetics for facial rejuvenation and dermatological treatments in the clinic. However, there is a tradeoff between therapeutic effectiveness and possible adverse effects caused by this agent for cosmetologists. To optimize the cosmetic efficacy with minimal concentration, we proposed a chemical permeation enhancer (CPE) azone to synergistically work with SA on human skin in vivo. The optical properties of human skin after being treated with SA alone and SA combined with azone (SA@azone) were successively investigated by diffuse reflectance spectroscopy (DRS) and optical coherence tomography (OCT). Our results revealed that as the SA concentration increased, the light reflectance decreased and the absorption increased. We also found that SA@azone exhibited a synergistic effect on enhancing light penetration and OCT imaging depth. We demonstrated that the combination of DRS and OCT techniques could be used as a noninvasive, rapid and accurate measurement method to monitor the subtle changes of skin tissue after treatment with FCPA and CPE. The approach will greatly benefit the development of clinical cosmetic surgery, dermatosis diagnosis and therapeutic effect inspection in related biomedical studies.

  7. Non-respiratory tuberculosis with Mycobacterium tuberculosis after penetrating lesions of the skin : five case histories

    NARCIS (Netherlands)

    de Jong, JW; van Altena, R

    2000-01-01

    Tuberculosis is primarily transmitted from person to person via the respiratory route. We describe five cases of patients who developed tuberculosis at the site of a skin injury: three after being treated repeatedly with local corticosteroids via intramuscular injections, and two who cut themselves

  8. Effect of Simultaneous Administration of Dihydroxyacetone on the Diffusion of Lawsone Through Various In Vitro Skin Models.

    Science.gov (United States)

    Munt, Daniel J; Grana, Anne; Hulce, Martin; Fusaro, Ramon M; Dash, Alekha K

    2015-12-01

    Unprotected sunlight exposure is a risk factor for a variety of cutaneous cancers. Topically used dihydroxyacetone (DHA) creates, via Maillard reaction, chemically fixed keratin sunscreen in the stratum corneum with significant protection against UVA/Soret radiation. When used in conjunction with naphthoquinones a naphthoquinone-modified DHA Maillard reaction is produced that provides protection across the UVB/UVA/Soret spectra lasting up to 1 week, resisting sweating and contact removal. The aim of this study was to examine a simplified version of this formulation for effect on UV transmission and to determine if penetration levels merit toxicity concerns. Permeability was demonstrated for freshly prepared DHA (30 mg/mL) and lawsone (0.035 mg/mL) alone and in combination using a side-by-side diffusion apparatus at 37°C over 48 h across shed snake skin and dermatomed pig skin. These samples were then examined for effectiveness and safety. Concentrations were determined by HPLC and UPLC monitored from 250-500 nm. Lawsone flux significantly decreased across pig skin (20.8 (± 4.8) and 0.09 (± 0.1) mg/cm(2) h without and with DHA, respectively) but did not change across shed snake skin in the presence of DHA. Significantly reduced lawsone concentration was noted in donor chambers of combined solutions. Damage was not observed in any skins. Darker coloration with greater UV absorbance was observed in skins exposed to the combined solution versus individual solutions. This study confirmed that combined DHA and lawsone provided effective blocking of ultraviolet light through products bound in keratinized tissue. DHA permeation levels in pig skin suggest further in vitro and in vivo study is required to determine the safety of this system.

  9. Human in vitro skin organ culture as a model system for evaluating DNA repair.

    Science.gov (United States)

    Liu, Hannah; Tuchinda, Papapit; Fishelevich, Rita; Harberts, Erin; Gaspari, Anthony A

    2014-06-01

    UV-exposures result in accumulation of genetic lesions that facilitate the development of skin cancer. Numerous pharmacologic agents are currently under development to both inhibit formation of DNA lesions and enhance repair. Drugs must be evaluated in vitro, currently performed in cell culture systems, before being tested on humans. Current systems do not account for the architecture and diverse cellularity of intact human skin. To establish a novel, functionally viable, and reproducible in vitro skin organ culture system for studying the effects of various pharmacologic agents on DNA repair. Human skin was obtained from neonatal foreskins. Intact skin punches derived from foreskins were cultured in vitro prior to exposure to UV-irradiation, and evaluated for DNA-damage using a DNA dot blot. Serial skin biopsies were obtained from patients with actinic keratoses treated with topical imiquimod. Expression of immune-stimulating and DNA repair genes was evaluated in ex vivo and in vitro samples. DNA dot blots revealed active repair of UV induced lesions in our in vitro skin organ culture. The photo-protective effect of sunscreen was detected, while imiquimod treatment did not enhance DNA repair in vitro. The DNA repair molecules XPA and XPF were up-regulated in the skin of imiquimod treated patients with actinic keratoses and imiquimod treated bone marrow-derived cell lines, but not keratinocytes. Our in vitro human skin organ culture model detected repair of UV-induced DNA lesions, and may be easily adapted to investigate various photo-protective drugs intended to prevent or treat skin cancer. Copyright © 2014 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.

  10. Permeation of chromium salts through human skin in vitro

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Fullerton, A; Avnstorp, C

    1992-01-01

    of the dichromate solution. Chromium skin levels increased with increasing concentrations of applied chromium salts up to 0.034 M Cr. The amount of chromium in recipient phase and skin layers increased with increasing pH when the applied solution contained potassium dichromate. This was ascribed to a decreased skin...... barrier function of the skin. The amount of chromium found in all skin layers after application of chromium chloride decreased with increasing pH due to lower solubility of the salt. The % of chromium found in the recipient phase as chromium(VI) increased with increasing total chromium concentration...

  11. The effects of chemical and physical penetration enhancers on the percutaneous permeation of lidocaine through equine skin

    Science.gov (United States)

    2014-01-01

    Background The effect of physical and chemical permeation enhancers on in vitro transdermal permeation of lidocaine was investigated in the horse. Therefore, the effect of six vehicles (phosphate-buffered saline (PBS), 50% ethanol, 50% propylene glycol, 50% isopropylalcohol, 50% isopropylalcohol/isopropylmyristate and 50% dimethylsulfoxide) was examined as well as the effect of microneedle pretreatment with different needle lengths on transdermal drug delivery of lidocaine. The skin was obtained from the thorax of six Warmblood horses and was stored up to two weeks at - 20°C. Franz-type diffusion cells were used to study the transdermal permeation through split skin (600 μm thickness). The amount of lidocaine in the receptor fluid was determined by UV–VIS high-performance liquid chromatography. Results All investigated vehicle supplementations diminished the transdermal flux of lidocaine through equine skin in comparison to pure PBS except dimethylsulfoxide, which resulted in comparable permeation rates to PBS. The maximum flux (Jmax) was 1.6-1.8 fold lower for lidocaine applied in 50% ethanol, propylene glycol, isopropylalcohol and isopropylalcohol/isopropylmyristate. A significant higher Jmax of lidocaine was observed when lidocaine was applied in PBS onto microneedle pretreated skin with similar permeation rates in both needle lengths. After 6 hours, 1.7 fold higher recovery rates were observed in the microneedle pretreated skin samples than in the untreated control samples. The lagtimes were reduced to 20–50% in the microneedle pretreated skin samples. Conclusion Microneedles represent a promising tool for transdermal lidocaine application in the horse with a rapid systemic bioavailability. PMID:24950611

  12. Topical drug delivery by a polymeric nanosphere gel: Formulation optimization and in vitro and in vivo skin distribution studies.

    Science.gov (United States)

    Batheja, Priya; Sheihet, Larisa; Kohn, Joachim; Singer, Adam J; Michniak-Kohn, Bozena

    2011-01-20

    Tyrosine-derived nanospheres have demonstrated potential as effective carriers for the topical delivery of lipophilic molecules. In this investigation, a gel formulation containing nanospheres was developed for effective skin application and enhanced permeation. Carbopol and HPMC hydrophilic gels were evaluated for dispersion of these nanospheres. Sparingly water soluble diclofenac sodium (DS) and lipophilic Nile Red were used as model compounds. DS was used to determine the optimum polymer type, viscosity and release properties of the gel while fluorescent Nile Red was used in in vitro and in vivo skin distribution studies. In addition, the effect of a penetration enhancer, Azone, on the skin delivery was investigated. Dispersion of Nile Red-loaded nanospheres in 1% w/v HPMC gel produced a uniform and stable dispersion with suitable rheological properties for topical application, without any short-term cellular toxicity or tissue irritation. In vitro permeation studies using human cadaver skin revealed that the deposition of Nile Red via the nanosphere gel in the upper and lower dermis was 1.4 and 1.8 fold higher, respectively, than the amount of Nile Red deposited via an aqueous nanosphere formulation. In vivo, the HPMC gel containing Nile Red-loaded nanospheres significantly enhanced (1.4 fold) the permeation of Nile Red to the porcine stratum corneum/epidermis compared to the aqueous Nile Red-loaded nanospheres. An additional increase (1.4 fold) of Nile Red deposition in porcine stratum corneum/epidermis was achieved by incorporation of Azone (0.2M) into the nanosphere gel formulation. Therefore, tyrosine-derived nanospheres dispersed in gels offer promise for the topical delivery of lipophilic drugs and personal care agents to skin for treatment of cancers, psoriasis, eczema, and microbial infections. Copyright © 2010 Elsevier B.V. All rights reserved.

  13. Permeação cutânea in vitro do ácido kójico Kojic acid in vitro percutaneous penetration study

    Directory of Open Access Journals (Sweden)

    Mayumi Eliza Otsuka Sato

    2007-06-01

    .4, both containing kojic acid (2%. The kojic acid percutaneous penetration and retention was quantified by a spectrophotometric in the ultraviolet method. The in vitro study showed that the absorption of kojic acid from the formulation presented kinetic of pseudo 1st order (in the break from 1 to 8 hours and consequently smaller stream through the natural membrane (skin from the ear of pig and bigger cutaneous retention, while the kojic acid from the buffered solution pH 7.4 presented kinetic of zero order and consequently bigger stream and smaller retention. This result indicated that the formulation developed can be used as a vehicle for kojic acid, having in mind that the target tissue is the skin.

  14. Correlation of sperm penetration assay score with polyspermy rate in in-vitro fertilization.

    Science.gov (United States)

    Aoki, Vincent W; Peterson, C Matthew; Parker-Jones, Kirtly; Hatasaka, Harry H; Gibson, Mark; Huang, Ivan; Carrell, Douglas T

    2005-02-09

    BACKGROUND: The sperm penetration assay (SPA) is used to predict the fertilizing capacity of sperm. Thus, some programs rely on SPA scores to formulate insemination plans in conjunction with in-vitro fertilization (IVF) cycles. The purpose of this study was to evaluate if a relationship exists between SPA scores and polyspermy rates during conventional IVF cycles. METHODS: A total of 1350 consecutive IVF patients using conventional IVF insemination were evaluated in the study. Oocytes were inseminated three hours post-retrieval by the addition of 150,000 to 300,000 progressively motile sperm. Approximately 18 hours after insemination, the oocytes were evaluated for fertilization by the visualization of pronuclei. The presence of three or more pronuclei was indicative of polyspermy. Polyspermy rates, fertilization success, embryo quality, and pregnancy rates were analyzed retrospectively to evaluate their relationship with SPA score, count, motility, number of progressively motile sperm inseminated, oocyte pre-insemination incubation time, patient age, and diagnosis. RESULTS: A significant positive relationship was observed between SPA score and polyspermy rate (rs = 0.10, p polyspermy rates than those with abnormal SPA scores (6.3% +/- 1.5% vs. 2.0% +/- 0.7%, p polyspermy rates and IVF fertilization percentage. Additionally, there is a slight increase in clinical pregnancy rates, and embryo implantation rates with increased SPA. Furthermore, there is a slight decrease in spontaneous abortions rates related to increased SPA.

  15. In vitro dermal absorption of decabromodiphenyl ethane in rat and human skin

    Data.gov (United States)

    U.S. Environmental Protection Agency — In vitro dermal absorption of decabromodiphenyl ethane in rat and human skin. This dataset is associated with the following publication: Knudsen, G., J.M. Sanders,...

  16. Determination of in vitro usnic acid delivery into porcine skin using a HPLC method.

    Science.gov (United States)

    Serafini, Mairim Russo; Detoni, Cassia Britto; Guterres, Sílvia Stanisçuaski; da Silva, Gabriel Francisco; de Souza Araújo, Adriano Antunes

    2015-01-01

    Usnic acid, a lichen metabolite, has been proposed as a potential topical treatment for microbial skin lesions, burn wounds as well as a sunscreen. An isocratic HPLC method was validated according to FDA's Guidance for Industry: Bioanalytical Method Validation to determine skin penetration and permeation of usnic acid. The penetration and permeation of usnic acid was evaluated using Franz cells and porcine skin. The method was valid according to selectivity, linearity, precision, accuracy and stability. Usnic acid was quantified in the skin surface (6.13 µg cm(2)), stratum corneum (34.4 µg cm(2)), viable epidermis (5.6 µg cm(2)), dermis (28.2 µg cm(2)) and receptor compartment (3.2 µg cm(2)). These results help us to understand the penetration profile of usnic acid and plan topical therapeutic approaches as well as new topical delivery systems to modulate this penetration profile. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  17. Porcine ear skin as a biological substrate for in vitro testing of sunscreen performance.

    Science.gov (United States)

    Sohn, Myriam; Korn, Verena; Imanidis, Georgios

    2015-01-01

    The purpose of the study was to examine the use of skin from porcine ears as a biological substrate for in vitro testing of sunscreens in order to overcome the shortcomings of the presently used polymethylmethacrylate (PMMA) plates that generally fail to yield a satisfactory correlation between sun protection factors (SPF) in vitro and in vivo. Trypsin-separated stratum corneum and heat-separated epidermis provided UV-transparent substrates that were laid on quartz or on PMMA plates. These were used to determine surface roughness by chromatic confocal imaging and to measure SPF in vitro of 2 sunscreens by diffuse transmission spectroscopy. The recovered skin layers showed a lower roughness than full-thickness skin but yielded SPF in vitro values that more accurately reflected the SPF determined in vivo by a validated procedure than PMMA plates, although the latter had in part roughness values identical to those of intact skin. Combination of skin tissue with a high roughness PMMA plate also provided accurate SPF in vitro. Besides roughness, the improved affinity of the sunscreen to the skin substrate compared to PMMA plates may explain the better in vitro prediction of SPF achieved with the use of a biological substrate. © 2014 S. Karger AG, Basel.

  18. Combined use of borneol or menthol with labrasol promotes penetration of baicalin through rabbit cornea in vitro.

    Science.gov (United States)

    Huang, Lei; Bai, Jianhai; Yang, Hongbin; Liu, Jingjing; Cui, Hao

    2015-01-01

    The permeability of most drugs through the eyes is very limited, so finding safe and effective penetration enhancers is of high importance in current ophthalmology research. In this paper, we use a new approach that integrates Chinese and Western medicine to improve the corneal permeability of baicalin, a water- and fat-insoluble target drug, in vitro. Rabbits were divided into three groups. The first group was dosed with borneol (0.05%, 0.1%). menthol (0.1%, 0.2%), or Labrasol (1%, 2%) individually, the second was dosed with a combination of Labrasol with either borneol or menthol, and the third group received a control treatment. Compared with the control treatment, borneol, menthol, or Labrasol alone clearly improved the permeability of baicalin in vitro. Furthermore, the penetrating effects were significantly increased by combining the application of Labrasol with menthol or borneol. Among the various combined penetration enhancers, 0.1% borneol with 2% Labrasol achieved the best apparent permeability, approximately 16.35 times that of the control. Additionally, the calculation of corneal hydration level and the Draize test demonstrated the safety of these penetration enhancers to the rabbit corneas in vivo. This study confirms that the combined use of borneol or menthol, compounds both derived from Chinese herbs, with Labrasol can improve the corneal permeability of water- and fat-insoluble drugs.

  19. Staphylococcus aureus penetrate the interkeratinocyte spaces created by skin-infiltrating neutrophils in a mouse model of impetigo.

    Science.gov (United States)

    Imanishi, Ichiro; Hattori, Shinpei; Hisatsune, Junzo; Ide, Kaori; Sugai, Motoyuki; Nishifuji, Koji

    2017-02-01

    Impetigo is a bacterial skin disease characterized by intraepidermal neutrophilic pustules. Previous studies have demonstrated that exfoliative toxin producing staphylococci are isolated in the cutaneous lesions of human and canine impetigo. However, the mechanisms of intraepidermal splitting in impetigo remain poorly understood. To determine how staphylococci penetrate the living epidermis and create intraepidermal pustules in vivo using a mouse model of impetigo. Three Staphylococcus aureus strains harbouring the etb gene and three et gene negative strains were epicutaneously inoculated onto tape-stripped mouse skin. The skin samples were subjected to time course histopathological and immunofluorescence analyses to detect intraepidermal neutrophils and infiltrating staphylococci. To determine the role of neutrophils on intraepidermal bacterial invasion, cyclophosphamide (CPA) was injected intraperitoneally into the mice to cause leucopenia before the inoculation of etb gene positive strains. In mice inoculated with etb gene positive S. aureus, intraepidermal pustules resembling impetigo were detected as early as 4 h post-inoculation (hpi). Neutrophils in the epidermis were detected from 4 hpi, whereas intraepidermal staphylococci was detected from 6 hpi. The dimensions of the intraepidermal clefts created in mice inoculated with etb gene positive strains at 6 hpi were significantly larger than those in mice inoculated with et gene negative strains. In CPA treated mice, staphylococci or neutrophils were not detected in the deep epidermis until 6 hpi. Our findings indicate that intraepidermal neutrophils play an important role in S. aureus invasion into the living epidermis in a mouse model of impetigo. © 2016 ESVD and ACVD.

  20. Ultra-small lipid nanoparticles promote the penetration of coenzyme Q10 in skin cells and counteract oxidative stress.

    Science.gov (United States)

    Lohan, Silke B; Bauersachs, Sonja; Ahlberg, Sebastian; Baisaeng, Nuttakorn; Keck, Cornelia M; Müller, Rainer H; Witte, Ellen; Wolk, Kerstin; Hackbarth, Steffen; Röder, Beate; Lademann, Jürgen; Meinke, Martina C

    2015-01-01

    UV irradiation leads to the formation of reactive oxygen species (ROS). An imbalance between the antioxidant system and ROS can lead to cell damage, premature skin aging or skin cancer. To counteract these processes, antioxidants such as coenzyme Q10 (CoQ10) are contained in many cosmetics. To improve and optimize cell/tissue penetration properties of the lipophilic CoQ10, ultra-small lipid nanoparticles (usNLC) were developed. The antioxidant effectiveness of CoQ10-loaded usNLC compared to conventional nanocarriers was investigated in the human keratinocyte cell line HaCaT. Using confocal laser scanning microscopy investigations of the carriers additionally loaded with nile red showed a clear uptake into cells and their distribution within the cytoplasm. By use of the XTT cell viability test, CoQ10 concentrations of 10-50 μg/ml were shown to be non-toxic, and the antioxidant potential of 10 μg/ml CoQ10 loaded usNLC in the HaCaT cells was analyzed via electron paramagnetic resonance spectroscopy after cellular exposure to UVA (1J/cm(2)) and UVB (18 mJ/cm(2)) irradiation. In comparison with the CoQ10-loaded conventional carriers, usNLC-CoQ10 demonstrated the strongest reduction of the radical formation; reaching up to 23% compared to control cells without nanocarrier treatment. Therefore, usNLC-CoQ10 are very suitable to increase the antioxidant potential of skin. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. A preliminary comparison of total skin electron treatment techniques to demonstrate the application of a mid-torso phantom for measurement of dose penetration.

    Science.gov (United States)

    Baugh, G; Al-Alawi, T; Fletcher, C L; Mills, J A; Grieve, R J

    2011-12-01

    In the UK, the treatment of patients with mycosis fungoides using total skin electron (TSE) beam therapy is undertaken using a number of different irradiation techniques. As part of a review of these techniques, a comparative set of measurements would be useful to determine how the techniques differ in terms of dose distribution. A dose penetration intercomparison method that could be used as part of such a study is presented here. The dose penetrations for six treatment techniques currently or recently used in four centres in the UK were measured. The variation of dose with skin depth was measured in a WT1 solid water mid-torso phantom. The phantom is portable and suitable to be used in all the techniques. It is designed to hold four small radiochromic film dosemeters to investigate the variation in dose around the mid-torso. For each treatment technique, the phantom was irradiated using the clinical set-up. The phantom performed well and was able to measure dose penetration and the uniformity of penetration for several treatment techniques. These preliminary results demonstrate that there is some variation in dose distribution between different TSE treatment techniques and that the phantom could be used in a more comprehensive intercomparison. The results are not intended to demonstrate comprehensively the range of penetration that can be achieved in clinical practice as, for one of the treatment techniques, the penetration is customised for the extent of the disease.

  2. Development of the isolated perfused porcine skin flap for in vitro studies of percutaneous absorption pharmacokinetics and cutaneous biotransformation

    Energy Technology Data Exchange (ETDEWEB)

    Carver, M.P.

    1988-01-01

    The isolated perfused porcine skin flap (IPPSF) has proven to be a valuable in vitro tool for studying the physiology and biochemistry of skin and for identifying biochemical and histological markers of direct cutaneous toxicity. The present experiments were undertaken for two purposes: (1) to develop a pharmacokinetic model, based on dermal penetration in the IPPSF, which is predictive of percutaneous absorption in vivo, and (2) to examine cutaneous biotransformation of the important agricultural poison parathion (P). Dosing solutions of {sup 14}C-radiolabelled compounds representing 3 chemical classes-organic acid/base (benzoic acid (B), caffeine (C)), organophosphate (OP) pesticides, and steroid hormones, were applied topically in ethanol at 40 {mu}m cm{sup {minus}2}, both in vivo and on the IPPSF. A 3-compartment pharmacokinetic model describing mass transfer from the surface (C{sub 1}), diffusion through epidermis and dermis (C{sub 2}), and transfer into the perfusate (C{sub 3}), was developed based on flux through the IPPSF from 0-8 hr. Model simulations were predictive of percutaneous absorption in vivo for the OP's and steroids. Modification of the basic 3-compartment model to account for fast and slow tissue-release processes (B) and for flux-dependent perfusage flow increases (C), provided excellent in vivo-in vitro correlation over all 7 compounds.

  3. Impact of Humidity on In Vitro Human Skin Permeation Experiments for Predicting In Vivo Permeability.

    Science.gov (United States)

    Ishida, Masahiro; Takeuchi, Hiroyuki; Endo, Hiromi; Yamaguchi, Jun-Ichi

    2015-12-01

    In vitro skin permeation studies have been commonly conducted to predict in vivo permeability for the development of transdermal therapeutic systems (TTSs). We clarified the impact of humidity on in vitro human skin permeation of two TTSs having different breathability and then elucidated the predictability of in vivo permeability based on in vitro experimental data. Nicotinell(®) TTS(®) 20 and Frandol(®) tape 40mg were used as model TTSs in this study. The in vitro human skin permeation experiments were conducted under humidity levels similar to those used in clinical trials (approximately 50%) as well as under higher humidity levels (approximately 95%). The skin permeability values of drugs at 95% humidity were higher than those at 50% humidity. The time profiles of the human plasma concentrations after TTS application fitted well with the clinical data when predicted based on the in vitro permeation parameters at 50% humidity. On the other hand, those profiles predicted based on the parameters at 95% humidity were overestimated. The impact of humidity was higher for the more breathable TTS; Frandol(®) tape 40mg. These results show that in vitro human skin permeation experiments should be investigated under realistic clinical humidity levels especially for breathable TTSs. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  4. Topical administration of tetrasodium-meso-tetraphenyl-porphinesulfonate (TPPS): correlation between drug penetration and depth of necrosis in skin of nude mice following red light irradiation.

    Science.gov (United States)

    Marchesini, R; Melloni, E; Bottiroli, G; Andreola, S; Fava, G; Mella, M; Savi, G; Zunino, F

    1987-02-28

    The main side effect in photodynamic therapy is photosensitization of the patient's skin following systemic administration of the photosensitizing agent. In the case of superficial lesions, this problem can be avoided by topically applying the drug: in this way a local treatment can be performed. We tested the photosensitizing properties of a 2% solution of TPPS (tetrasodium-tetraphenylporphinesulfonate) in a vehicle containing a penetration enhancer, Azone, on skin of nude mice. An aliquot of 0.1 ml/cm2 of the solution was painted on the skin overlying an s.c. implanted NMU-1 tumor. Subsequently, animals were sacrificed at different times after application. Fluorescence microscopy revealed that TPPS penetration depth was related to time elapsed after application and to painting modalities. Solution penetration was enhanced by wiping with ether immediately before painting. Irradiation at 80 mW/cm2 for 20 min with a dye laser emitting at 640 nm, 4 h after TPPS application, produced necrosis of the upper skin layers, up to 0.2 mm in depth. These findings suggest that topical TPPS administration, followed by laser irradiation, may be a suitable treatment modality for skin lesions involving epithelial layers, even though several aspects of this metodology need further investigation.

  5. Comparison of ATR-FTIR spectra of porcine vaginal and buccal mucosa with ear skin and penetration analysis of drug and vehicle components into pig ear.

    Science.gov (United States)

    Schwarz, Julia C; Pagitsch, Elisabeth; Valenta, Claudia

    2013-12-18

    In the present study, porcine buccal and vaginal mucosae were successfully characterised by ATR-FTIR for the first time and compared to porcine ear skin. By analysing typical bands of the spectra, the structure of proteins and the lipid matrix were elucidated. According to the body site, differences in membrane permeability were detected when analysing the CH2-stretching and -scissoring vibrations. The results indicated a higher permeability for porcine vaginal and buccal tissue compared to skin. Furthermore, the influence of a lecithin-based microemulsion on the barrier properties of the above mentioned tissues was investigated by ATR-FTIR; the results revealed structural changes in all tissues. In addition, the ATR-FTIR technique was employed to semi-quantitatively analyse compounds directly on skin. To this end, tape stripping experiments were performed with a deuterated liposomal drug delivery system containing the model drug flufenamic acid. While the amount of penetrated deuterated liposomes was determined directly on skin samples by ATR-FTIR, the drug amount was analysed by HPLC after extraction of the tape strips since higher sensitivity was achieved in this fashion. Thus, it was possible to monitor the skin penetration of drug and vehicle simultaneously. Interestingly, the results indicated an independent drug penetration after release from the liposomal carrier system. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. In-vitro percutaneous absorption of losartan potassium in human skin and prediction of human skin permeability

    Directory of Open Access Journals (Sweden)

    Petkar K.C.

    2007-05-01

    Full Text Available This study describes the feasibility of transdermal controlled administration of Losartan potassium (LP across human cadaver skin. Study also defines the influence of capsaicin, sex and site of application on permeation characteristics and determined an appropriate animal model for human skin permeability. The permeation of LP of various formulations was studied using Keshary-Chein diffusion cell. Optimized controlled formulation (without capsaicin released 42.17% (±1.85 of LP in 12 hr whereas treatment formulation (with capsaicin 0.028 % w/v released 48.94% (±1.71 of LP with significant difference on null hypothesis. Influence of sex showed statistically significant difference for permeation of LP through male and female rats, as well as male and female mice across both the abdominal and dorsal sides of the skin (p<0.05. Similarly statistically significant differences were noted for permeation of LP across male and female mice abdomen-dorsal, but not for male rat abdomen-dorsal and female rat abdomen-dorsal. Furthermore, in-vitro permeation of LP across human skin was compared with the permeation across rat and mice skins. Male rat and male mice dorsal skin was found to have closer permeability characteristics to human than other skin membranes, but the Factor of Difference values were < 3 for all membranes which were used suggesting the membranes are good models for human skin permeability. In conclusion simple transdermal adhesive patches formulations incorporating high molecular weight of LP can deliver a dose in-vivo and proposed model skin membranes can be utilized for future pharmacokineic and toxicokinetic studies as well as metabolism studies of LP

  7. Skin penetration of infective hookworm larvae. II. The path of migration of infective larvae of Ancylostoma braziliense in the metacarpal foot pads of dogs.

    Science.gov (United States)

    Vetter, J C; van der Linden, M E

    1977-11-10

    The hairless metacarpal foot pads of six hookworm-free puppies were exposed to infective larvae of Ancylostoma braziliense. Serial sections of the biopts stained with Harris' haematoxylin and eosin showed that the infective larvae are able to penetrate the toughest region of canine skin. Pores of eccrine sweat glands did not seem to constitute sites of entry and no larvae were detected in these glands. Larvae were only observed in the epidermis. The histopathology of the infected skin of the foot pads of the puppies was similar to that in human skin with "creeping eruption" as described by Fülleborn (1927). The biopts appeared to consist of hairy skin as well. In the unexposed adjacent hairy skin of the foot pads, larvae were also observed. They were found in the epidermis, hair follicle systems and dermis, suggesting that the migration from the epidermis into deeper tissue depends on the presence of the hair follicle systems.

  8. Ethosomes for skin delivery of ropivacaine: preparation, characterization and ex vivo penetration properties.

    Science.gov (United States)

    Zhai, Yingjie; Xu, Rui; Wang, Yi; Liu, Jiyong; Wang, Zimin; Zhai, Guangxi

    2015-01-01

    Ropivacaine, a novel long-acting local anesthetic, has been proved to own superior advantage. However, Naropin® Injection, the applied form in clinic, can cause patient non-convenience. The purpose of this study was to formulate ropivacaine (RPV) in ethosomes and evaluate the potential of ethosome formulation in delivering RPV transdermally. The RPV-loaded ethosomes were prepared with thin-film dispersion technique and the formulation was characterized in terms of size, zeta potential, differential scanning calorimetry (DSC) analysis and X-ray diffraction (XRD) study. The results showed that the optimized RPV-ethosomes displayed a typical lipid bilayer structure with a narrow size distribution of 73.86 ± 2.40 nm and drug loading of 8.27 ± 0.37%, EE of 68.92 ± 0.29%. The results of DSC and XRD study indicated that RPV was in amorphous state when encapsulated into ethosomes. Furthermore, the results of ex vivo permeation study proved that RPV-ethosomes could promote the permeability in a high-efficient, rapid way (349.0 ± 11.5 μg cm(-2) at 12 h and 178.8 ± 7.1 μg cm(-2) at 0.5 h). The outcomes of histopathology study forecasted that the interaction between ethosomes and skin could loosen the tight conjugation of corneocyte layers and weaken the permeation barrier. In conclusion, RPV-ethosomes could be a promising delivery system to encapsulate RPV and deliver RPV for transdermal administration.

  9. The Pharmaceutical Device Prisma® Skin Promotes in Vitro Angiogenesis through Endothelial to Mesenchymal Transition during Skin Wound Healing

    Directory of Open Access Journals (Sweden)

    Raffaella Belvedere

    2017-07-01

    Full Text Available Glycosaminoglycans are polysaccharides of the extracellular matrix supporting skin wound closure. Mesoglycan is a mixture of glycosaminoglycans such as chondroitin-, dermatan-, heparan-sulfate and heparin and is the main component of Prisma® Skin, a pharmaceutical device developed by Mediolanum Farmaceutici S.p.a. Here, we show the in vitro effects of this device in the new vessels formation by endothelial cells, since angiogenesis represents a key moment in wound healing. We found a strong increase of migration and invasion rates of these cells treated with mesoglycan and Prisma® Skin which mediate the activation of the pathway triggered by CD44 receptor. Furthermore, endothelial cells form longer capillary-like structures with a great number of branches, in the presence of the same treatments. Thus, the device, thanks to the mesoglycan, leads the cells to the Endothelial-to-Mesenchymal Transition, suggesting the switch to a fibroblast-like phenotype, as shown by immunofluorescence assays. Finally, we found that mesoglycan and Prisma® Skin inhibit inflammatory reactions such as nitric oxide secretion and NF-κB nuclear translocation in endothelial cells and Tumor Necrosis Factor-α production by macrophages. In conclusion, based on our data, we suggest that Prisma® Skin may be able to accelerate angiogenesis in skin wound healing, and regulate inflammation avoiding chronic, thus pathological, responses.

  10. Evaluation of the in vitro skin permeation of antiviral drugs from penciclovir 1% cream and acyclovir 5% cream used to treat herpes simplex virus infection

    Directory of Open Access Journals (Sweden)

    Bader Marlene

    2009-04-01

    Full Text Available Abstract Background Herpes simplex virus infection (HSV is a common and ubiquitous infection of the skin which causes mucocutaneous lesions called cold sores (herpes labialis or fever blisters. It is estimated that approximately 80% of the population worldwide are carriers of the Herpes simplex virus, approximately 40% suffer from recurrent recurrent infections. This study evaluates the in vitro skin permeation and penetration of penciclovir and acyclovir from commercialized creams for the treatment of herpes labialis (cold sores, using non viable excised human abdominal skin samples, which were exposed to 5 mg/cm2 of acyclovir 5% cream or penciclovir 1% cream. Methods After 24 h of cream application, excess cream was washed off and layers of stratum corneum were removed by successive tape stripping. Amounts of active ingredients having penetrated through the skin were measured, as well as the amounts in the washed-off cream, in skin strips and creams remaining in the skin. Molecular modelling was used to evaluate physico-chemical differences between the drugs. Western blot analysis enabled to determine whether the marker of basal cells keratin 5 could be detected in the various tape strips. Results Application of penciclovir 1% cream yielded higher concentration of drug in the deeper layers of the epidermis as well as a higher drug flux through the skin. Molecular modelling showed two higher hydrophobic moieties for acyclovir. Presence of the basal cell marker keratin 5 was underscored in the deeper tape strips from the skin, giving evidence that both drugs can reach their target cells. Conclusion Penciclovir 1% cream has the tendency to facilitate the diffusion of the drug through the stratum corneum into the deeper epidermis layers, in which it could reach the target basal cells at effective therapeutical concentration. The small difference in the surface properties between both molecules might also contribute to favour the passage of

  11. Mechanisms of skin adherence, penetration and tissue necrosis production by Haemophilus ducreyi, the causative agent of chancroid.

    Science.gov (United States)

    Abeck, D; Korting, H C

    1992-01-01

    Haemophilus ducreyi (H. ducreyi) strains, representing both reference strains and low-passage isolates, were investigated in terms of surface structures and enzymatic equipment. The interaction of these factors with host tissue was analysed using new in vitro- and in vivo-models. By electron microscopy studies there was no evidence of an extracellular capsule or surface appendages such as pili or flagella. Interaction of all isolates tested with the lectin Phaseolus vulgaris suggests N-acetyl-D-glucosamine units as common structural features of H. ducreyi cell envelope polysaccharide. In attachment to epithelial cells more than one hemagglutinin might be implicated as different haemagglutination patterns could be observed whereby the activity was not heat-labile, but was abolished by formaldehyde. Hydrophobic interactions might be of importance as well as strains showed a wide range of reactions from hydrophobic to hydrophilic, low hydrophobicity being more marked with the older strains. No elaboration of degradative enzymes based on the measurement of enzymatic activity using insoluble dye-protein complexes could be detected in case of H. ducreyi, using Azocoll and Remazol Brilliantblue hide powder for detection of proteolytic activity and elastinorcein for detection of elastase activity. In vitro studies using human keratinocytes and Vero cells did not show any morphological changes when incubated with H. ducreyi culture filtrates. In vivo studies with a new mouse model for H. ducreyi infection could confirm the results of the in vitro studies. Mere contact to undamaged skin both of whole cell organisms, live or heat-killed, and of culture filtrates did not lead to any reaction or even damage of mouse skin. However, when the outer epidermal layer was overcome by intradermal injection of shaved mice ulcers developed. Tissue necrosis production was not bound to live organisms as dead ones showed the same effect. There is great evidence that this tissue necrosis is

  12. From the structure of the skin barrier and dermal formulations to in vitro transport models for skin absorption: skin research in the Netherlands and in Germany.

    Science.gov (United States)

    Windbergs, M; Hansen, S; Schroeter, A; Schaefer, U F; Lehr, C-M; Bouwstra, J

    2013-01-01

    This review presents an overview of German and Dutch research institutions and their studies in the field of skin drug delivery and adjacent topics. In the Netherlands, the involved research groups are mainly localized in Leiden, whereas in Germany the skin research institutions are spread over the whole country. The scientific studies in the Netherlands focus on the in-depth analysis of human skin composition and its individual components as well as on the development and characterization of dermal drug delivery systems ranging from liquid crystalline systems and vesicles up to microneedles with an emphasis on examining the interactions of these drug delivery systems with the human skin in vitro and in vivo. In Germany, the individual areas of research span from in-depth investigations on various drug delivery systems intended for skin application and the development of novel in vitro models for skin absorption testing up to in vivo studies focusing on the biological performance of topically applied actives. Furthermore, sophisticated analytical techniques are applied for the elucidation of skin assembly and transport processes. In addition, experimentally derived data are correlated with advanced computational modelling. Even though the individual research topics in the Netherlands and Germany are quite diverse, the exchange of knowledge and interdisciplinary collaborations between the two neighbouring countries were and are still frequently made. In this context, the review aims at highlighting crosslinks between the different institutions and individual persons to complete the picture. For each institution, the principal investigators and their studies are presented and the upcoming young scientists are introduced as an outlook for the field. This review does not claim completeness, but is rather intended to give a general overview of Dutch and German research in the field of skin drug delivery and adjacent topics. © 2013 S. Karger AG, Basel.

  13. In Vitro Nail Penetration of Tavaborole Topical Solution, 5%, Through Nail Polish on Ex Vivo Human Fingernails.

    Science.gov (United States)

    Vlahovic, Tracey; MPharm, Tejal Merchant; Chanda, Sanjay; Zane, Lee T; Coronado, Dina

    2015-07-01

    Onychomycosis is a common infection of the toenails that causes nail thickening and discoloration. The physical appearance of the infected nail can diminish self-image and negatively impact quality of life. Patients may use nail polish to mask the appearance of infected nails. To evaluate the in vitro nail penetration properties of tavaborole topical solution, 5%, through nail polish using ex vivo, non-diseased human fingernails. In study 1, tavaborole penetration was evaluated over 20 days of dosing using the Franz finite dose technique and modified Franz diffusion cells. Nails received either 1 coat of over-the-counter (OTC) typical polish or were left unpolished (controls). In study 2, tavaborole penetration was measured over 14 days of dosing using the finite dose technique and vertical diffusion cells. Nails were polished with either 4 coats or 1 coat of salon typical polish or with 2 coats or 1 coat of OTC typical polish, or they were left unpolished. In study 1, the mean ± standard deviation (SD) cumulative tavaborole penetration at day 21 was numerically higher, though not statistically significant, through polished nails (3,526 ± 1,433 μg/cm(2))vs unpolished nails (2,661 ± 1,319 μg/cm(2)).In study 2, the mean cumulative tavaborole penetration was also numerically higher (statistical significance not assessed) through all nails that received polish vs unpolished nails. At day 15, mean ± SD cumulative tavaborole nail penetration was 1,179 ± 554 μg/cm(2) through 4 coats of salon typical polish, 1,227 ± 974 μg/cm(2) through 1 coat of salon typical polish, 1,493 ± 1,322 μg/cm(2) through 2 coats of OTC typical polish, 1,428 ± 841 μg/cm(2) through 1 coat of OTC typical polish, and 566 ± 318 μg/cm(2) through unpolished nails. Results from these in vitro studies demonstrated that tavaborole penetrated through human nails with up to 4 layers of nail polish.

  14. Intraocular pressure-lowering effect of oral paracetamol and its in vitro corneal penetration properties

    Directory of Open Access Journals (Sweden)

    Mohamed N

    2013-01-01

    Full Text Available Nabiel Mohamed, David MeyerDivision of Ophthalmology, Faculty of Medicine and Health Sciences, University of Stellenbosch, Cape Town, South AfricaBackground: Several studies have confirmed the ability of cannabinoids to reduce intraocular pressure. Experimental data recently demonstrated unequivocally that the analgesic effect of paracetamol is due to its indirect action on cannabinoid receptors. The question then arises as to whether paracetamol can reduce intraocular pressure via its effect on intraocular cannabinoid receptors.Methods: A 2-week, prospective, randomized, controlled, single-center, parallel-group pilot study was carried out to determine the efficacy and safety of paracetamol 1 g orally administered every 6 hours in adult patients with primary or secondary open angle glaucoma as compared with topical levobunolol 0.5% twice a day. Patient well-being was closely monitored throughout the study and focused on hepatic safety in accordance with Drug-Induced Liver Injury Network criteria. The in vitro diffusion kinetics of acetaminophen in a phosphate-buffered solution in rabbit and human corneas was also investigated, with the view to a topical application.Results: Eighteen adult patients were enrolled in the study, with nine in the topical levobunolol group and nine in the oral paracetamol group. In the levobunolol group, the mean reduction in intraocular pressure at day 7 was 7.5 mmHg (P < 0.008 and at day 14 was 9.1 mmHg (P < 0.005, from a mean baseline intraocular pressure of 29.6 mmHg. The corresponding figures for the paracetamol group were 8.8 mmHg (P < 0.0004 at day 7 and 6.5 mmHg (P < 0.004 at day 14, from a mean baseline intraocular pressure of 29.4 mmHg. Both study regimens were well tolerated. No serious treatment-related adverse events were reported in either of the treatment groups. Liver function tests, systolic/diastolic blood pressure, or heart rate remained unchanged in both groups during the 2 weeks of the study. In

  15. In vitro skin absorption tests of three types of parabens using a Franz diffusion cell.

    Science.gov (United States)

    Seo, Ji-Eun; Kim, Sungkyoon; Kim, Bae-Hwan

    2017-05-01

    The objective of this study was to evaluate the permeation of paraben derivatives - methylparaben (MP), propylparaben (PP), and butylparaben (BP) - in hairless mouse full skin and human cadaver epidermis using a Franz diffusion cell method, which is proposed as a reliable alternative method to an skin absorption test. Parabens, esterified hydroxybenzoic acid compounds, are widely used as preservatives in food, cosmetics, and pharmaceutical products. The skin permeation rate showed dose dependency, and the hairless mouse full skin showed a higher flux value than human cadaver epidermis. Among the permeability coefficient (Kp) values of three parabens, MP showed a higher Kp value than PP or BP. Hence, according to the definitions of Marzulli et al., parabens would be classified as "moderate" penetrants.

  16. Penetration of silver nanoparticles into porcine skin ex vivo using fluorescence lifetime imaging microscopy, Raman microscopy, and surface-enhanced Raman scattering microscopy.

    Science.gov (United States)

    Zhu, Yongjian; Choe, Chun-Sik; Ahlberg, Sebastian; Meinke, Martina C; Alexiev, Ulrike; Lademann, Juergen; Darvin, Maxim E

    2015-05-01

    In order to investigate the penetration depth of silver nanoparticles (Ag NPs) inside the skin, porcine ears treated with Ag NPs are measured by two-photon tomography with a fluorescence lifetime imaging microscopy (TPT-FLIM) technique, confocal Raman microscopy (CRM), and surface-enhanced Raman scattering (SERS) microscopy. Ag NPs are coated with poly-N-vinylpyrrolidone and dispersed in pure water solutions. After the application of Ag NPs, porcine ears are stored in the incubator for 24 h at a temperature of 37°C. The TPT-FLIM measurement results show a dramatic decrease of the Ag NPs' signal intensity from the skin surface to a depth of 4 μm. Below 4 μm, the Ag NPs' signal continues to decline, having completely disappeared at 12 to 14 μm depth. CRM shows that the penetration depth of Ag NPs is 11.1 ± 2.1 μm. The penetration depth measured with a highly sensitive SERS microscopy reaches 15.6 ± 8.3 μm. Several results obtained with SERS show that the penetration depth of Ag NPs can exceed the stratum corneum (SC) thickness, which can be explained by both penetration of trace amounts of Ag NPs through the SC barrier and by the measurements inside the hair follicle, which cannot be excluded in the experiment.

  17. Penetration of silver nanoparticles into porcine skin ex vivo using fluorescence lifetime imaging microscopy, Raman microscopy, and surface-enhanced Raman scattering microscopy

    Science.gov (United States)

    Zhu, Yongjian; Choe, Chun-Sik; Ahlberg, Sebastian; Meinke, Martina C.; Alexiev, Ulrike; Lademann, Juergen; Darvin, Maxim E.

    2015-05-01

    In order to investigate the penetration depth of silver nanoparticles (Ag NPs) inside the skin, porcine ears treated with Ag NPs are measured by two-photon tomography with a fluorescence lifetime imaging microscopy (TPT-FLIM) technique, confocal Raman microscopy (CRM), and surface-enhanced Raman scattering (SERS) microscopy. Ag NPs are coated with poly-N-vinylpyrrolidone and dispersed in pure water solutions. After the application of Ag NPs, porcine ears are stored in the incubator for 24 h at a temperature of 37°C. The TPT-FLIM measurement results show a dramatic decrease of the Ag NPs' signal intensity from the skin surface to a depth of 4 μm. Below 4 μm, the Ag NPs' signal continues to decline, having completely disappeared at 12 to 14 μm depth. CRM shows that the penetration depth of Ag NPs is 11.1±2.1 μm. The penetration depth measured with a highly sensitive SERS microscopy reaches 15.6±8.3 μm. Several results obtained with SERS show that the penetration depth of Ag NPs can exceed the stratum corneum (SC) thickness, which can be explained by both penetration of trace amounts of Ag NPs through the SC barrier and by the measurements inside the hair follicle, which cannot be excluded in the experiment.

  18. Topical formulations containing finasteride. Part I: in vitro permeation/penetration study and in vivo pharmacokinetics in hairless rat.

    Science.gov (United States)

    Monti, Daniela; Tampucci, Silvia; Burgalassi, Susi; Chetoni, Patrizia; Lenzi, Carla; Pirone, Andrea; Mailland, Federico

    2014-08-01

    In hair follicle (Hf) cells, the type-2 5-α-reductase enzyme, implicated in androgenetic alopecia, is selectively inhibited by finasteride (FNS). Because an effective topical formulation to deliver FNS to Hf is currently unavailable, this investigation aimed at evaluating in vitro FNS skin permeation and retention through and into hairless rat and human abdominal skin. Four hydroxypropyl chitosan (HPCH)-based formulations (P-08-012, P-08-016, P-08-063, and P-08-064) and one anhydrous formulation without HPCH (P-10-008) were tested. The pharmacokinetics in plasma and skin after application of P-08-016 or P-10-008 on dorsal rat skin with single and repeated doses was investigated. P-08-016 performed the best in driving FNS to the reticular dermis without producing a high transdermal flux. Neither the in vivo single nor the repeated dose experiments produced plasma levels of FNS and no differences were found between formulations concerning skin retention. No increase in the amount of drug retained in the skin was obtained with the repeated dose experiment. In conclusion, the HPCH-based formulation P-08-016 might represent an alternative to systemic therapy for its ability to promote a cutaneous depot of FNS in the region of hair bulbs, minimizing systemic absorption even after repeated treatments. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  19. Comparison of Sealer Penetration by Using Different Irrigation Techniques - An In-vitro Study.

    Science.gov (United States)

    Ismail, Prabu Mahin Syed; Ahamed, Siddiq; Sabiha, P B; Sekhar, M Chandra; Moosani, Gopikrishna; Reddy, S Nagalakshmi; Reddy, N Upendranatha; Sumanthi, C H

    2016-12-01

    The main goal of root canal treatment is to eliminate the microorganisms particularly in the apical third area and to prevent re-infection. In order to achieve these goals the instrumentation must be combined with adequate irrigation. To compare sealer penetration by using different irrigation techniques i.e., apical negative pressure irrigation, Passive Ultrasonic Irrigation (PUI) and combination of apical negative pressure irrigation and PUI. A total of 48 single rooted maxillary central incisors were taken. Access cavity was prepared and biomechanical preparation was done. The samples were randomly assigned into three experimental groups based on the final irrigation technique used. Group I: Apical negative pressure (Endovac); Group II: PUI; Group III: Combination of apical negative pressure and PUI. All the samples were obturated using AH plus sealer and the sections were observed under confocal laser scanning microscope to evaluate the percentage and maximum depth of sealer penetration at 1mm, 3mm and 5mm levels. Statistical analysis was done by using two way ANOVA and Tukey's post-hoc test to compare the percentage and maximum depth of sealer penetration. Combination group resulted in better sealer penetration at 1mm and 3mm from the working length than the Endovac and PUI group. However, the Endovac group showed significantly better sealer penetration at 1mm from the working length when compared with PUI. There was no significant difference in sealer penetration at 5mm level between PUI and combination group. Combination group was the only group to achieve better sealer penetration at 1mm and 3mm levels from the working length.

  20. Prediction of formulation effects on dermal absorption of topically applied ectoparasiticides dosed in vitro on canine and porcine skin using a mixture-adjusted quantitative structure permeability relationship.

    Science.gov (United States)

    Riviere, J E; Brooks, J D; Collard, W T; Deng, J; de Rose, G; Mahabir, S P; Merritt, D A; Marchiondo, A A

    2014-10-01

    Topical application of ectoparasiticides for flea and tick control is a major focus for product development in animal health. The objective of this work was to develop a quantitative structure permeability relationship (QSPeR) model sensitive to formulation effects for predicting absorption and skin deposition of five topically applied drugs administered in six vehicle combinations to porcine and canine skin in vitro. Saturated solutions (20 μL) of (14) C-labeled demiditraz, fipronil, permethrin, imidacloprid, or sisapronil were administered in single or binary (50:50 v/v) combinations of water, ethanol, and transcutol (6 formulations, n = 4-5 replicates per treatment) nonoccluded to 0.64 cm(2) disks of dermatomed pig or dog skin mounted in flow-through diffusion cells. Perfusate flux over 24 h and skin deposition at termination were determined. Permeability (logKp), absorption, and penetration endpoints were modeled using a four-term Abrahams and Martin (hydrogen-bond donor acidity and basicity, dipolarity/polarizability, and excess molar refractivity) linear free energy QSPeR equation with a mixture factor added to compensate for formulation ingredient interactions. Goodness of fit was judged by r(2) , cross-validation coefficient, coefficients (q(2) s), and Williams Plot to visualize the applicability domain. Formulation composition was the primary determinant of permeation. Compounds generally penetrated dog skin better than porcine skin. The vast majority of permeated penetrant was deposited within the dosed skin relative to transdermal flux, an attribute for ectoparasiticides. The best QSPeR logKp model for pig skin permeation (r(2) = 0.86, q(2) s = 0.85) included log octanol/water partition coefficient as the mixture factor, while for dogs (r(2) = 0.91, q(2) s = 0.90), it was log water solubility. These studies clearly showed that the permeation of topical ectoparasiticides could be well predicted using QSPeR models that account for both the physical

  1. Fabrication, in-vitro characterization, and enhanced in-vivo evaluation of carbopol-based nanoemulsion gel of apigenin for UV-induced skin carcinoma.

    Science.gov (United States)

    Jangdey, Manmohan S; Gupta, Anshita; Saraf, Swarnlata

    2017-11-01

    The aim of this study was to develop a potential novel formulation of carbopol-based nanoemulsion gel containing apigenin using tamarind gum emulsifier which was having the smallest droplet size, the highest drug content, and a good physical stability for Skin delivery. Apigenin loaded nanoemulsion was prepared by high speed homogenization method and they were characterized with respect to morphology, zeta potential, differential scanning calorimeter study, and penetration studies. In-vitro release studies and skin permeation of apigenin loaded nanoemulsion by goat abdominal skin was determined using Franz diffusion cell and confocal laser scanning microscope (CLSM). The cytotoxicity of the reported formulation was evaluated in HaCaT Cells (A) and A431 cells (B) by MTT assay. The nanoemulsion formulation showed droplet size, polydispersity index, and zeta potential of 183.31 nm, 0.532, and 31.9 mV, respectively. The nanoemulsions were characterized by TEM demonstrated spherical droplets and FTIR to ensure the compatibility among its ingredients. CLSM showed uniform fluorescence intensity across the entire depth of skin in nanocarriers treatment, indicating high penetrability of nanoemulsion gel through goatskin. The nanoemulsion gel showed toxicity on melanoma (A341) in a concentration range of 0.4-2.0 mg/ml, but less toxicity toward HaCaT cells. The carbopol-based nanoemulsion gel formulation of apigenin possesses better penetrability across goatskin as compared to marketed formulation. Hence, the study postulates that the novel nanoemulsion gel of apigenin can be proved fruitful for the treatment of skin cancer in near future.

  2. Enhanced in vitro and in vivo skin deposition of apigenin delivered using ethosomes.

    Science.gov (United States)

    Shen, Li-Na; Zhang, Yong-Tai; Wang, Qin; Xu, Ling; Feng, Nian-Ping

    2014-01-02

    The aim of this study was to develop and evaluate a novel topical delivery system for apigenin by using ethosomes. An optimal apigenin-loaded ethosome formulation was identified by means of uniform design experiments. Skin deposition and transdermal flux of apigenin loaded in ethosomes, liposomes, and deformable liposomes were compared in vitro and in vivo. The efficiency of apigenin encapsulation increased with an increase in the amount of phospholipids in ethosome formulations. Moreover, skin deposition and transdermal flux of apigenin improved with an increase in the levels of phospholipids (Lipoid S 75) and short-chain alcohols (propylene glycol and ethanol), but decreased with an increase in the ratio of propylene glycol to ethanol. Profiles of skin deposition versus time for ethosomes varied markedly between in vivo and in vitro studies compared with those of liposomes or deformable liposomes. Optimized ethosomes showed superior skin targeting both in vitro and in vivo. Moreover, they had the strongest effect on reduction of cyclooxygenase-2 levels in mouse skin inflammation induced by ultraviolet B (UVB) light. Therefore, apigenin-loaded ethosomes represent a promising therapeutic approach for the treatment of UVB-induced skin inflammation. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Examining the Impact of Skin Lighteners In Vitro

    Directory of Open Access Journals (Sweden)

    James V. Gruber

    2013-01-01

    Full Text Available Three cosmetically important skin lightening agents, hydroquinone (HQ, kojic acid (KA, and niacinamide (NA, consume the bulk of successful skin lightening ingredients in cosmetic applications. However, the mechanisms by which these ingredients work are still unclear. In this study, melanocytes and keratinocytes were treated with high, nontoxic doses of HQ, KA, and NA and the cells were examined by human microarrays and protein assays for several important targets including cytotoxicity, melanin expression, tyrosinase gene (TYR and protein expression, melanocortin-1 receptor (MC1R gene and protein expression, cytochrome c oxidase-1 (COX1 gene and protein expression, and ferritin (FTH1 gene and protein expression. It was found that all the skin lighteners examined showed marked increases in TYR, COX1, and FTH1 gene and protein expression, but not in MC1R expression in melanocytes. Upregulation of COX1 and FTH1 genes and proteins was common across both cell lines, melanocytes and keratinocytes. The results of the tyrosinase expression were somewhat unexpected. The role of iron in the expression of melanin is somewhat unexplored, but common and strong upregulation of ferritin protein in both types of cells due to the treatments suggests that iron plays a more pivotal role in melanin synthesis than previously anticipated.

  4. Examining the Impact of Skin Lighteners In Vitro

    Science.gov (United States)

    Gruber, James V.; Holtz, Robert

    2013-01-01

    Three cosmetically important skin lightening agents, hydroquinone (HQ), kojic acid (KA), and niacinamide (NA), consume the bulk of successful skin lightening ingredients in cosmetic applications. However, the mechanisms by which these ingredients work are still unclear. In this study, melanocytes and keratinocytes were treated with high, nontoxic doses of HQ, KA, and NA and the cells were examined by human microarrays and protein assays for several important targets including cytotoxicity, melanin expression, tyrosinase gene (TYR) and protein expression, melanocortin-1 receptor (MC1R) gene and protein expression, cytochrome c oxidase-1 (COX1) gene and protein expression, and ferritin (FTH1) gene and protein expression. It was found that all the skin lighteners examined showed marked increases in TYR, COX1, and FTH1 gene and protein expression, but not in MC1R expression in melanocytes. Upregulation of COX1 and FTH1 genes and proteins was common across both cell lines, melanocytes and keratinocytes. The results of the tyrosinase expression were somewhat unexpected. The role of iron in the expression of melanin is somewhat unexplored, but common and strong upregulation of ferritin protein in both types of cells due to the treatments suggests that iron plays a more pivotal role in melanin synthesis than previously anticipated. PMID:23738040

  5. Nanocrystalline titanium dioxide and magnesium oxide in vitro dermal absorption in human skin.

    Science.gov (United States)

    van der Merwe, Deon; Tawde, Snehal; Pickrell, John A; Erickson, Larry E

    2009-01-01

    The dermal absorption potential of a nanocrystalline magnesium oxide (MgO) and titanium dioxide (TiO(2)) mixture in dermatomed human skin was assessed in vitro using Bronaugh-type flow-through diffusion cells. Nanocrystalline material was applied to the skin surface at a dose rate of 50 mg/cm(2) as a dry powder, as a water suspension, and as a water/surfactant (sodium lauryl sulfate) suspension, for 8 hours. Dermal absorption of nanocrystalline MgO and TiO(2) through human skin with intact, functional stratum corneum was not detectable under the conditions of this experiment.

  6. IN VITRO AND IN VIVO EVALUATION OF SILVER NANOPARTICLES PENETRATION THROUGH HUMAN SKIN

    OpenAIRE

    Bianco, Carlotta

    2015-01-01

    La cute è uno degli organi più estesi del corpo umano e gioca un importante ruolo nella regolazione dell’idratazione corporea, ha proprietà sensoriali, funzioni strutturali e agisce come prima barriera contro gli agenti esterni (Blank et al, 1984). Può costituire un’importante via di uptake per molte sostanze. L’assorbimento percutaneo è stato oggetto di studio in numerosi lavori fin dallo scorso secolo, ma ha recentemente riscosso nuovo interesse a causa dell’ascesa del “mondo delle nanopart...

  7. Optical clearing of porcine skin tissue in vitro studied by Raman microspectroscopy

    Science.gov (United States)

    Huang, Deqiu; Zhang, Wen; Zhong, Huiqing; Xiong, Honglian; Guo, Xi; Guo, Zhouyi

    2012-01-01

    In present work, we studied the effect of optical clearing on porcine skin in vitro with glycerol by Raman microspectroscopy, denoted as RM, at various time intervals of 0, 15, 30, 45, 60, and 75 min respectively. The results showed that the addition of glycerol significantly improved the depth of RM measurement, and enhanced the recovery of skin tissue Raman spectra that were not overlapped with the glycerol Raman spectra over time. Moreover, it was found that the Raman signals resembled the native spectrum of the molecules in porcine skin with a negligible frequency shift. Furthermore, we evaluated the extent of optical clearing in porcine skin by utilizing various concentrations of 40%, 60%, and 80% glycerol solution. The results demonstrated that with the increase of concentration of glycerol, the optical clearing of porcine skin was much improved.

  8. Standardization procedure for the in vitro skin permeation of anticholinergics

    NARCIS (Netherlands)

    Bosman, I.J; Ensing, K; de Zeeuw, R.A

    1998-01-01

    The permeation of seven anticholinergics was studied in vitro on pig epidermal membranes, using static Franz diffusion cells. The donor solution consisted of isotonic phosphate-buffered saline, pH 7.4 with ethanol, propylene glycol and Azone(R). Tritium-labelled dexetimide was added as an internal

  9. AN IN-VITRO EVALUATION OF FLUORESCEIN PENETRATION INTO NATURAL ROOT SURFACE CARIOUS LESIONS

    NARCIS (Netherlands)

    VANDERVEEN, MH; TENBOSCH, JJ

    1993-01-01

    In order to develop a method for detection and quantification of initial root surface carious lesions, the use of fluorescein sodium as a fluorescent dye is evaluated. The penetration depth of fluorescein sodium into human roots containing natural carious lesions was measured on approximately

  10. Characterization of Temperature Profiles in Skin and Transdermal Delivery System When Exposed to Temperature Gradients In Vivo and In Vitro.

    Science.gov (United States)

    Zhang, Qian; Murawsky, Michael; LaCount, Terri; Hao, Jinsong; Kasting, Gerald B; Newman, Bryan; Ghosh, Priyanka; Raney, Sam G; Li, S Kevin

    2017-07-01

    Performance of a transdermal delivery system (TDS) can be affected by exposure to elevated temperature, which can lead to unintended safety issues. This study investigated TDS and skin temperatures and their relationship in vivo, characterized the effective thermal resistance of skin, and identified the in vitro diffusion cell conditions that would correlate with in vivo observations. Experiments were performed in humans and in Franz diffusion cells with human cadaver skin to record skin and TDS temperatures at room temperature and with exposure to a heat flux. Skin temperatures were regulated with two methods: a heating lamp in vivo and in vitro, or thermostatic control of the receiver chamber in vitro. In vivo basal skin temperatures beneath TDS at different anatomical sites were not statistically different. The maximum tolerable skin surface temperature was approximately 42-43°C in vivo. The temperature difference between skin surface and TDS surface increased with increasing temperature, or with increasing TDS thermal resistance in vivo and in vitro. Based on the effective thermal resistance of skin in vivo and in vitro, the heating lamp method is an adequate in vitro method. However, the in vitro-in vivo correlation of temperature could be affected by the thermal boundary layer in the receiver chamber.

  11. Gene delivery into human skin in vitro using biphasic lipid vesicles.

    Science.gov (United States)

    Foldvari, Marianna; Kumar, Praveen; King, Martin; Batta, Ravinder; Michel, Deborah; Badea, Ildiko; Wloch, Mary

    2006-01-01

    Topical gene delivery to the skin shows great potential for painless, non-invasive administration of novel vaccines and therapeutic agents. The challenge is to develop a pharmaceutically acceptable system that can deliver suitable amounts of plasmid DNA to produce the desired level of response. The purpose of this study was to quantitatively assess DNA delivery by a novel lipid-based biphasic delivery system into the viable layers of excised human skin. Biphasic lipid vesicle formulations, incorporating plasmid DNA were evaluated in vitro in flow-through diffusion cells. Fifty mg DNA formulation containing 10 microg DNA was applied to full-thickness human breast skin for 24 hours. Residual formulation was removed and the skin was washed with PBS, then tape-stripped, followed by DNase treatment to remove surface bound DNA. Skin samples were homogenised and digested overnight with Proteinase K. The resulting supernatant was used as a template for quantitative PCR. Three formulations yielded a significant degree of dermal absorption compared to the controls. Formulation 26-3-2-DNA indicated that approximately 1x10(9) copies of plasmid were absorbed per cm2 skin. Other formulations resulted in 5x10(6) copies/cm2 skin (17C3-1-DNA) and 5x10(8) copies/cm2 skin (26-3-1-DNA). Biphasic vesicles delivered significant quantities of plasmid DNA into the 'viable' layers of human skin in vitro. The successful delivery of this large (approximately 4,400 kDa) charged molecule through intact stratum corneum represents a major advance in transdermal macromolecule delivery.

  12. Transdermal iontophoresis of the dopamine agonist 5-OH-DPAT in human skin in vitro

    NARCIS (Netherlands)

    Nugroho, AK; Li, L; Dijkstra, D; Wikstrom, H; Danhof, M; Bouwstra, JA

    2005-01-01

    The feasibility of transdermal iontophoretic delivery of a potent dopamine agonist 5-OH-DPAT was studied in vitro in side by side diffusion cells across human stratum corneum (HSC) and dermatomed human skin (DHS) according to the following protocol: 6 h of passive diffusion, 9 h of iontophoresis and

  13. Visualization studies of human skin in vitro/in vivo under the influence of an electrical field

    NARCIS (Netherlands)

    Fatouros, N.E.; Groenink, H.W.M.; Graaff, de A.M.; Aelst, van A.C.; Koerten, H.K.; Bouwstra, J.A.

    2006-01-01

    The aim of this study was to investigate the local changes in the ultrastructure of human skin after iontophoresis, using cryo-scanning, transmission and freeze fracture electron microscopy in human skin in vitro and in vivo. Human dermatomed skin was subjected to passive diffusion for 6 hours

  14. Development of a Full-Thickness Human Skin Equivalent In Vitro Model Derived from TERT-Immortalized Keratinocytes and Fibroblasts

    NARCIS (Netherlands)

    Reijnders, C.M.A.; van Lier, A.; Roffel, S.; Kramer, D.; Scheper, R.J.; Gibbs, S.

    2015-01-01

    Currently, human skin equivalents (HSEs) used for in vitro assays (e.g., for wound healing) make use of primary human skin cells. Limitations of primary keratinocytes and fibroblasts include availability of donor skin and donor variation. The use of physiologically relevant cell lines could solve

  15. [Study on preparation of testosterone undecanoate ethosomes and its in vitro transdermal penetration].

    Science.gov (United States)

    Meng, Shu; Yang, Li-Qun; Ma, Li-Ying; Guo, Jing; Li, Miao; Yang, Dan

    2013-05-01

    Ethosomes, as a new vector for transdermal drug delivery, could obviously improve the transdermal penetration of drugs. In this study, we prepared testosterone undecanoate ethosomes, with TU ethosomes as the basic remedy, to determine its appearance, particle size, entrapment efficiency (EE) and membrane fluidity. Meanwhile, a transdermal test was conducted in mice, in order to determine the permeability characteristics of ethosomes as a vector for transdermal drug delivery, and compare transdermal behaviors of TU ethosomes, liposomes and their ethanol solutions.

  16. Transdermal kinetics of a mercurous chloride beauty cream: an in vitro human skin analysis.

    Science.gov (United States)

    Palmer, R B; Godwin, D A; McKinney, P E

    2000-01-01

    Crema de Belleza-Manning is a popular mercurous chloride-containing beauty cream used to smooth and lighten the complexion and treat acne. Hundreds of people in the Southwestern US border states have been identified with elevated (>20 microg/L) urine mercury levels believed to be secondary to using this cream. The kinetic characteristics of percutaneous mercury absorption are incompletely defined. The objective of this study was to determine the transdermal kinetics of two formulations of mercurous chloride from a beauty cream in an in vitro human skin model. A proprietary formulation and an aqueous formulation of the beauty cream were studied using modified Franz diffusion cells. Mercury content in the skin samples and the underlying diffusion buffer was determined using atomic absorption spectrophotometry. A rapid initial increase in mercury content both in the skin and the buffer was noted for both formulations. Mercury concentrations in the aqueous samples were significantly (p < 0.05) higher in both the skin and the diffusion buffer compared to parallel samples containing glycerol. Mercury was readily absorbed through the skin in this in vitro human skin model. The aqueous preparation had a markedly increased rate and extent of mercury absorption relative to the proprietary formulation.

  17. In vitro techniques to assess the proficiency of skin care cosmetic formulations.

    Science.gov (United States)

    Roy, Amit; Sahu, Ram Kumar; Matlam, Munglu; Deshmukh, Vinay Kumar; Dwivedi, Jaya; Jha, Arvind Kumar

    2013-07-01

    Cosmetics comprising either natural or synthetic components are used almost regularly and universally in different forms to enhance the beauty. The utmost disclosure of human membrane to sunlight and environmental pollution results in the exhibition of free radical, that react with deoxyribonucleic acid, proteins and fatty acids, causation oxidative destruction dysfunction of the antioxidant system. In skin, the formation of reactive oxygen species leads to skin diseases, predominantly cutaneous malignancies, immunosuppression, wrinkles, aging, etc., The human organism fosters a barrier practice against the destructive action of free radicals, comprising mostly of vitamins, carotenoids and enzymes. Cosmetic products are the best option to reduce skin disorders such as hyper pigmentation, skin aging, skin wrinkling and rough skin texture, etc., Hence in this review, we conferred various in vitro methods that are used for the development of novel cosmetic formulation. There is an expanding fascinate employing in vitro techniques because they are less time consuming, more cost-effective and lessen the participation of human volunteers.

  18. In vitro skin decontamination of the organophosphorus pesticide Paraoxon with nanometric cerium oxide CeO2.

    Science.gov (United States)

    Salerno, Alicia; Devers, Thierry; Bolzinger, Marie-Alexandrine; Pelletier, Jocelyne; Josse, Denis; Briançon, Stéphanie

    2017-04-01

    Organophosphorus compounds (OP), which mainly penetrate via the percutaneous pathway, represent a threat for both military and civilians. Body surface decontamination is vital to prevent victims poisoning. The development of a cost-effective formulation, which could be efficient and easy to handle in case of mass contamination, is therefore crucial. Metal oxides nanoparticles, due their large surface areas and the large amount of highly reactive sites, present high reactivity towards OP. First, this study aimed at evaluating the reaction of CeO 2 nanoparticles, synthetized by microwave path and calcined at 500 or 600 °C, with Paraoxon (POX) in aqueous solution. Results showed that both nanoparticles degraded 60%-70% of POX. CeO 2 calcined at 500 °C, owing to its larger specific area, was the most effective. Moreover, the degradation was significantly increased under Ultra-Violet irradiation (initial degradation rate doubled). Then, skin decontamination was studied in vitro using the Franz cell method with pig-ear skin samples. CeO 2 powder and an aqueous suspension of CeO 2 (CeO 2 -W) were applied 1 h after POX exposure. The efficiency of decontamination, including removal and/or degradation of POX, was compared to Fuller's earth (FE) and RSDL lotion which are, currently, the most efficient systems for skin decontamination. CeO 2 -W and RSDL were the most efficient to remove POX from the skin surface and decrease skin absorption by 6.4 compared to the control not decontaminated. FE reduced significantly (twice) the absorbed fraction of POX, contrarily to CeO 2 powder. Considering only the degradation rate of POX, the products ranged in the order CeO 2  > RSDL > CeO 2 -W > FE (no degradation). This study showed that CeO 2 nanoparticles are a promising material for skin decontamination of OP if formulated as a dispersion able to remove POX like CeO 2 -W and to degrade it as CeO 2 powder. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  19. In vitro models for evaluating safety and efficacy of novel technologies for skin drug delivery.

    Science.gov (United States)

    Planz, Viktoria; Lehr, Claus-Michael; Windbergs, Maike

    2016-11-28

    For preclinical testing of novel therapeutics, predictive in vitro models of the human skin are required to assess efficacy, absorption and safety. Simple as well as more sophisticated three-dimensional organotypic models of the human skin emerged as versatile and powerful tools simulating healthy as well as diseased skin states. Besides addressing the demands of research and industry, such models serve as valid alternative to animal testing. Recently, the acceptance of several models by regulatory authorities corroborates their role as important building block for preclinical development. However, valid assessment of readout parameters derived from these models requires suitable analytical techniques. Standard analytical methods are mostly destructive and limited regarding in-depth investigation on molecular level. The combination of adequate in vitro models with modern non-invasive analytical modalities bears a great potential to address important skin drug delivery related questions. Topics of interest are for instance the assessment of repeated dosing effects and xenobiotic biotransformation, which cannot be analyzed by destructive techniques. This review provides a comprehensive overview of current in vitro skin models differing in functional complexity and mimicking healthy as well as diseased skin states. Further, benefits and limitations regarding analytical evaluation of efficacy, absorption and safety of novel drug carrier systems applied to such models are discussed along with a prospective view of anticipated future directions. In addition, emerging non-invasive imaging modalities are introduced and their significance and potential to advance current knowledge in the field of skin drug delivery is explored. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Natural oils as skin permeation enhancers for transdermal delivery of olanzapine: in vitro and in vivo evaluation.

    Science.gov (United States)

    Aggarwal, Geeta; Dhawan, Sanju; HariKumar, S L

    2012-03-01

    The feasibility of development of transdermal delivery system of olanzapine utilizing natural oils as permeation enhancers was investigated. Penetration enhancing potential of corn (maize) oil, groundnut oil and jojoba oil on in vitro permeation of olanzapine across rat skin was studied. The magnitude of flux enhancement factor with corn oil, groundnut oil and jojoba oil was 7.06, 5.31 and 1.9 respectively at 5mg/ml concentration in solvent system. On the basis of in vitro permeation studies, eudragit based matrix type transdermal patches of olanzapine were fabricated using optimized concentrations of natural oils as permeation enhancers. All transdermal patches were found to be uniform with respect to physical characteristics. The interaction studies carried out by comparing the results of ultraviolet, HPLC and FTIR analyses for the pure drug, polymers and mixture of drug and polymers indicated no chemical interaction between the drug and excipients. Corn oil containing unsaturated fatty acids was found to be promising natural permeation enhancer for transdermal delivery of olanzapine with greatest cumulative amount of drug permeated (1010.68 μg/cm²/h) up to 24 h and caused no skin irritation. The fabricated transdermal patches were found to be stable. The pharmacokinetic characteristics of the final optimized matrix patch (T2) were determined after transdermal application to rabbits. The calculated relative bioavailability of TDDS was 113.6 % as compared to oral administration of olanzapine. The therapeutic effectiveness of optimized transdermal system was confirmed by tranquillizing activity in rotarod and grip mice model.

  1. Percutaneous penetration of sodium lauryl sulphate is increased in uninvolved skin of patients with atopic dermatitis compared with control subjects

    NARCIS (Netherlands)

    Jakasa, I.; de Jongh, C. M.; Verberk, M. M.; Bos, J. D.; Kezić, S.

    2006-01-01

    BACKGROUND: Involved regions of the skin in patients with atopic dermatitis (AD) have been shown to have higher transepidermal water loss (TEWL), indicating a compromised skin barrier. Whether uninvolved skin also has diminished barrier characteristics is controversial. OBJECTIVES: To study the

  2. Examining the Genomic Influence of Skin Antioxidants In Vitro

    Directory of Open Access Journals (Sweden)

    James V. Gruber

    2010-01-01

    Full Text Available A series of well-known, purified antioxidants including: Resveratrol, Epigallocatechin Gallate (EGCG, Genistein, Rosavin, Puerarin, Chlorogenic Acid, Propolis and two newer unexplored isoflavonoids isolated from Maclura pomifera (Osage Orange including Pomiferin and Osajin, were applied to Normal Human Dermal Fibroblasts (NHDF and Normal Human Dermal Keratinocytes (NHEK for 24 hours. The resulting treated cells were then examined using human gene microarrays supplied by Agilent. These chips typically have somewhere on the order of 30,000 individual genes which are expressed in the human genome. For our study, this large list of genes was reduced to 205 principal genes thought to be important for skin and each individual ingredient was examined for its influence on the culled list of genes. Working on a hypothesis that there may be some common genes which are either upregulated or downregulated by all or most of these ingredients, a short list of genes for each cell line was developed. What appears to emerge from these studies is that several genes in the gene pool that was screened are influenced by most or all of the molecules of interest. Genes that appear to be upregulated in both cell lines by all the ingredients include: ACLY, AQP3, COX1, NOS3, and PLOD3. Genes that appear to be downregulated in both cell lines by all ingredients include only PGR.

  3. Examining the genomic influence of skin antioxidants in vitro.

    Science.gov (United States)

    Gruber, James V; Holtz, Robert

    2010-01-01

    A series of well-known, purified antioxidants including: Resveratrol, Epigallocatechin Gallate (EGCG), Genistein, Rosavin, Puerarin, Chlorogenic Acid, Propolis and two newer unexplored isoflavonoids isolated from Maclura pomifera (Osage Orange) including Pomiferin and Osajin, were applied to Normal Human Dermal Fibroblasts (NHDF) and Normal Human Dermal Keratinocytes (NHEK) for 24 hours. The resulting treated cells were then examined using human gene microarrays supplied by Agilent. These chips typically have somewhere on the order of 30,000 individual genes which are expressed in the human genome. For our study, this large list of genes was reduced to 205 principal genes thought to be important for skin and each individual ingredient was examined for its influence on the culled list of genes. Working on a hypothesis that there may be some common genes which are either upregulated or downregulated by all or most of these ingredients, a short list of genes for each cell line was developed. What appears to emerge from these studies is that several genes in the gene pool that was screened are influenced by most or all of the molecules of interest. Genes that appear to be upregulated in both cell lines by all the ingredients include: ACLY, AQP3, COX1, NOS3, and PLOD3. Genes that appear to be downregulated in both cell lines by all ingredients include only PGR.

  4. Development of an in vitro psoriatic skin model by tissue engineering.

    Science.gov (United States)

    Jean, Jessica; Lapointe, Marc; Soucy, Jacques; Pouliot, Roxane

    2009-01-01

    Psoriasis is a chronic skin disease characterized by a thickening and disorganization of the skin's protective barrier. This study aims to develop and characterize a novel in vitro psoriatic human skin model produced by tissue engineering. The self-assembly method, a tissue engineering approach based on the capacity of mesenchymal cells, such as fibroblasts, to create their own extracellular matrix in vitro, was used to create our substitutes. Manipulatable sheets of fibroblasts were superimposed creating a new dermis upon which keratinocytes are seeded, leading to a complete bilayered skin substitute. The characterization of the psoriatic substitutes was performed by macroscopic, histological and immunohistochemical analyses and contrasted to those constructed from healthy cells. Macroscopically, the psoriatic substitutes were more white and thicker than the healthy substitutes. The histological analysis of psoriatic substitutes stained with Masson's trichrome revealed a characteristic thickening of the epidermal layer seen in psoriatic skin in vivo. Immunohistochemical analysis of the psoriatic substitutes showed, among other things, an overexpression of involucrin and an underexpression of filaggrin and loricrin. These data suggest that the macroscopic, histological and immunohistochemical characteristics of psoriasis are partially retained in the substitutes, thus providing a good model to investigate the mechanisms of abnormal keratinocyte growth and to study cell-cell interactions.

  5. In vitro and in vivo germ line potential of stem cells derived from newborn mouse skin.

    Directory of Open Access Journals (Sweden)

    Paul W Dyce

    Full Text Available We previously reported that fetal porcine skin-derived stem cells were capable of differentiation into oocyte-like cells (OLCs. Here we report that newborn mice skin-derived stem cells are also capable of differentiating into early OLCs. Using stem cells from mice that are transgenic for Oct4 germline distal enhancer-GFP, germ cells resulting from their differentiation are expected to be GFP(+. After differentiation, some GFP(+ OLCs reached 40-45 µM and expressed oocyte markers. Flow cytometric analysis revealed that ∼ 0.3% of the freshly isolated skin cells were GFP(+. The GFP-positive cells increased to ∼ 7% after differentiation, suggesting that the GFP(+ cells could be of in vivo origin, but are more likely induced upon being cultured in vitro. To study the in vivo germ cell potential of skin-derived cells, they were aggregated with newborn ovarian cells, and transplanted under the kidney capsule of ovariectomized mice. GFP(+ oocytes were identified within a subpopulation of follicles in the resulting growth. Our finding that early oocytes can be differentiated from mice skin-derived cells in defined medium may offer a new in vitro model to study germ cell formation and oogenesis.

  6. Shed king cobra and cobra skins as model membranes for in-vitro nicotine permeation studies.

    Science.gov (United States)

    Pongjanyakul, Thaned; Prakongpan, Sompol; Panomsuk, Suwannee; Puttipipatkhachorn, Satit; Priprem, Aroonsri

    2002-10-01

    Shed king cobra skin (SKCS) and shed cobra skin (SCS) were investigated for use as barrier membranes, including some pre-hydration factors, for in-vitro nicotine permeation. Inter-specimen variations in nicotine fluxes using shed snake skin were compared with those using human epidermis. Nicotine in the form of 1% w/v aqueous buffer solution at pH 5 and transdermal patches (dose 14 mg day(-1)) were used. The nicotine fluxes across the shed snake skin were not significantly affected (P > 0.05) by temperature and duration of hydration pre-treatment. Scanning electron micrographs of SKCS and SCS revealed a remarkable difference in surface morphology, but the nicotine fluxes using both shed skins were not significantly different (P > 0.05). When compared with the results obtained using human epidermis, there were similarities in fluxes and permeation profiles of nicotine. Using nicotine solution, the nicotine permeation profiles of all membranes followed zero order kinetics. The amount of nicotine permeated provided good linearity with the square root of time over 24 h (R(2) > 0.98) when using nicotine patches. The nicotine fluxes using SKCS and SCS had less inter-specimen variation than those using human epidermis. The results suggest a potential use for SKCS or SCS as barrier membranes for in-vitro nicotine permeation studies.

  7. In vitro percutaneous penetration of acyclovir from solvent systems and Carbopol 971-P hydrogels: influence of propylene glycol.

    Science.gov (United States)

    Díez-Sales, O; Garrigues, T M; Herráez, J V; Belda, R; Martín-Villodre, A; Herráez, M

    2005-05-01

    The mechanism underlying propylene glycol (PG) effects on acyclovir (ACV) penetration through human epidermis were studied. Solvent systems and Carbopol gels containing increasing percentage of PG (from 0% to 70%, w/w) were used. Viscosity studies of both vehicles were carried out to characterise the influence of rheological behaviour. In solvent systems skin permeation values of ACV increase as the concentration of PG increase yielding a maximum enhancement ratio (ER = 10) for 70% PG. The release rate of ACV from gels was determined. Higuchi's model was used to estimate the apparent diffusion coefficient of the drug. These values show a decrease as the content of PG in the vehicle increases; this effect could be attributed to the increase of the viscosity in the diffusional pathway. When gels are used skin permeation values of ACV were smaller than those of the solvent systems. This could be attributed to the network structure created by the polymer that increases the length of the diffusional pathway. The maximum ER (= 6.8) was for Carbopol gel containing 50% PG. Therefore, these gels can be considered candidates for further research to confirm their usefulness as delivery systems for ACV topical formulations. Copyright 2005 Wiley-Liss, Inc

  8. Comparison of solvents for removing pesticides from skin using an in vitro porcine model.

    Science.gov (United States)

    Campbell, J L; Smith, M A; Eiteman, M A; Williams, P L; Boeniger, M F

    2000-01-01

    This study compared four solvents (1-propanol, polyethylene glycol [avg. MW 400], 10% Ivory Liquid and water, and D-TAM) for their ability to remove selected pesticides from an in vitro porcine skin model using a solvent-moistened wipe. Wipes were performed 90 min after pesticide was applied to the skin. The four pesticides selected (glyphosate, alachlor, methyl parathion, and trifluralin) were chosen because of their differences in water solubility. This study also determined whether pretreatment of skin with a solvent prior to pesticide application would either increase or decrease recovery of the pesticide. Recovery efficiencies for all solvents and pesticides were affected by the amount of contaminant on the skin. Although pesticide recoveries from all four solvents were similar (range: 45-57%), on average 1-propanol had significantly higher recoveries, followed by soap and water. There was no significant difference between polyethylene glycol, and D-TAM. When skin was pretreated with any of the four solvents before pesticide application, the recoveries of the more water soluble compounds, glyphosate and alachlor, decreased. When pretreatment with solvent preceded application of trifluralin, the pesticide with the lowest water solubility, recoveries increased. 1-Propanol or soap and water were more effective in removing pesticides from skin than polyethylene glycol or D-TAM, but the amount of pesticide recovered from skin was affected by the chemical characteristics of the pesticide (such as water solubility) and the amount of pesticide originally on the skin. This study provides information useful to the interpretation of skin wipe sample results collected in field studies.

  9. Effects of Sizes and Conformations of Fish-Scale Collagen Peptides on Facial Skin Qualities and Transdermal Penetration Efficiency

    Directory of Open Access Journals (Sweden)

    Huey-Jine Chai

    2010-01-01

    Full Text Available Fish-scale collagen peptides (FSCPs were prepared using a given combination of proteases to hydrolyze tilapia (Oreochromis sp. scales. FSCPs were determined to stimulate fibroblast cells proliferation and procollagen synthesis in a time- and dose-dependent manner. The transdermal penetration capabilities of the fractionationed FSCPs were evaluated using the Franz-type diffusion cell model. The heavier FSCPs, 3500 and 4500 Da, showed higher cumulative penetration capability as opposed to the lighter FSCPs, 2000 and 1300 Da. In addition, the heavier seemed to preserve favorable coiled structures comparing to the lighter that presents mainly as linear under confocal scanning laser microscopy. FSCPs, particularly the heavier, were concluded to efficiently penetrate stratum corneum to epidermis and dermis, activate fibroblasts, and accelerate collagen synthesis. The heavier outweighs the lighter in transdermal penetration likely as a result of preserving the given desired structure feature.

  10. Functionalization of mesoporous silica nanoparticles with a cell-penetrating peptide to target mammalian sperm in vitro.

    Science.gov (United States)

    Barkalina, Natalia; Jones, Celine; Townley, Helen; Coward, Kevin

    2015-05-01

    This study aimed to investigate the effects of actively targeting mesoporous silica nanoparticles (MSNPs) toward mammalian sperm with a cell-penetrating peptide (C105Y), with subsequent analysis of binding rates and nano-safety profiles. Boar sperm were exposed in vitro to C105Y-functionalized MSNPs or free C105Y, in a series of increasing doses for up to 2 h, followed by the evaluation of sperm motility, kinematic parameters, acrosome morphology, MSNP-sperm binding and cell fluorescence levels. C105Y-functionalized MSNPs preserved their biocompatibility with sperm, and exhibited an approximately fourfold increase in affinity toward gametes, compared with unmodified MSNPs, during the early stages of incubation. Our findings support the application of MSNPs and active targeting to sperm as valuable tools for reproductive biology.

  11. Chromium content in human skin after in vitro application of ordinary cement and ferrous-sulphate-reduced cement

    DEFF Research Database (Denmark)

    Fullerton, A; Gammelgaard, Bente; Avnstorp, C

    1993-01-01

    The amount of chromium found in human skin after in vitro application of cement suspensions on full-thickness human skin in diffusion cells was investigated. Cement suspensions made from ordinary Portland cement or Portland cement with the chromate reduced with added ferrous sulphate were used....... The cement suspensions were either applied on the skin surface under occlusion for 48 h or applied repeatedly every 24 h for 96 h. No statistically significant difference in chromium content of skin layers between skin exposed to ordinary Portland cement, skin exposed to cement with added ferrous sulphate...... and unexposed skin was observed, despite a more permeable skin barrier at the alkaline pH of the cement suspensions, i.e., pH 12.5. Increased chromium levels in epidermis and dermis were seen when ordinary Portland cement was applied as a suspension with added sodium sulphate (20%) on the skin surface for 96 h...

  12. Skin Toxicity Determined In vitro by Three-Dimensional, Native-State Histoculture

    Science.gov (United States)

    Li, Lingna; Margolis, Leonid B.; Hoffman, Robert M.

    1991-03-01

    We describe a gel-supported in vitro system for culturing skin samples in a three-dimensional native state. All the cell types of skin remain viable and maintain their native architecture for at least 10 days. The culture system is used for toxicity measurements by ascertaining cell viability using two fluorescent dyes: 2',7'-bis-(2-carboxyethyl)-5-(and -6)carboxyfluorescein acetoxymethyl ester, specific for living cells, and propidium iodide, specific for dead cells. Cell staining with the dyes is measured throughout the tissue block by confocal scanning fluorescence microscopy. The dose-response to three agents--ethanol, doxorubicin, and sodium hypochlorite--is shown and, in the case of sodium hypoclorite, compared to in vivo skin toxicity with a high correlation. We also demonstrate that the end point of [^3H]thymidine incorporation measured by histological autoradiography can be used to measure toxicity. Our results with the [^3H]thymidine end point demonstrate that the hair follicle cells are the most sensitive to doxorubicin. The native-state model for skin may be an effective replacement for animal systems and superior to the dispersed skin cell systems used previously. It can allow rapid, inexpensive measurements of the effect of manufactured products, drugs, and pollutants on skin.

  13. The effect of formulation on the penetration of coated and uncoated zinc oxide nanoparticles into the viable epidermis of human skin in vivo.

    Science.gov (United States)

    Leite-Silva, Vânia R; Le Lamer, Marina; Sanchez, Washington Y; Liu, David C; Sanchez, Washington H; Morrow, Isabel; Martin, Darren; Silva, Heron D T; Prow, Tarl W; Grice, Jeffrey E; Roberts, Michael S

    2013-06-01

    The use of nanoparticulate zinc oxide (ZnO-NP) in sunscreens and other cosmetic products has raised public health concerns. The two key issues are the extent of exposure to ZnO-NP and the likely hazard after the application of ZnO-NP in sunscreen and cosmetic products to humans in vivo. Our aims were to assess exposure by the extent of ZnO-NP penetration into the viable epidermis and hazard by changes in the viable epidermal redox state for a number of topical products. Of particular interest is the role of the particle coating, formulation used, and the presence of any enhancers. Multiphoton tomography with fluorescence lifetime imaging microscopy (MPT-FLIM) was used to simultaneously observe ZnO-NP penetration and potential metabolic changes within the viable epidermis of human volunteers after topical application of various ZnO-NP products. Coated and uncoated ZnO-NP remained in the superficial layers of the SC and in the skin furrows. We observed limited penetration of coated ZnO-NP dispersed in a water-in-oil emulsion formulation, which was predominantly localized adjacent to the skin furrow. However, the presence of ZnO-NP in the viable epidermis did not alter the metabolic state or morphology of the cells. In summary, our data suggest that some limited penetration of coated and uncoated ZnO-NP may occur into viable stratum granulosum epidermis adjacent to furrows, but that the extent is not sufficient to affect the redox state of those viable cells. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  14. In vitro skin permeation of sunscreen agents from O/W emulsions.

    Science.gov (United States)

    Montenegro, L; Carbone, C; Paolino, D; Drago, R; Stancampiano, A H; Puglisi, G

    2008-02-01

    The effects of different emulsifiers on the in vitro permeation through human skin of two sunscreen agents [octylmethoxycinnamate (OMC) and butylmethoxydibenzoylmethane (BMBM)] were investigated from O/W emulsions. The test formulations were prepared using the same oil and aqueous phase ingredients and the following emulsifier and coemulsifier systems: Emulgade SE((R)) (ceteareth-12 and ceteareth-20 and cetearyl alcohol and cetyl palmitate) and glycerylmonostearate (emulsion 1); Brij 72((R)) (steareth-2), Brij 721((R)) (steareth-21) and cetearyl alcohol (emulsion 2); Phytocream((R)) (potassium palmitoyl-hydrolysed wheat protein and glyceryl stearate and cetearyl alcohol) and glycerylmonostearate (emulsion 3); Montanov 68((R)) (cetearyl glucoside and cetearyl alcohol) (emulsion 4); Xalifin-15((R)) (C(15-20) acid PEG-8 ester) and cetearyl alcohol (emulsion 5). The cumulative amount of OMC that permeated in vitro through human skin after 22 h from the formulations being tested decreased in the order 3 > 1 congruent with 4 > 5 > 2 and was about nine-fold higher from emulsion 3 compared with that from emulsion 2. As regards BMBM, no significant difference was observed as regards its skin permeation from emulsions 1, 3, 4 and 5, whereas formulation 2 allowed significantly lower amounts of BMBM to permeate the skin. In vitro release experiments of OMC and BMBM from emulsions 1-6 through cellulose acetate membranes showed that only emulsions 4 and 5 provided pseudo-first-order release rates only for OMC. The results of this study suggest that the type of emulsifying systems used to prepare an O/W emulsion may strongly affect sunscreen skin permeation from these formulations. Therefore, the vehicle effects should be carefully considered in the formulation of sunscreen products.

  15. A simplified approach for estimating skin permeation parameters from in vitro finite dose absorption studies.

    Science.gov (United States)

    Lehman, Paul A

    2014-12-01

    Historically, percutaneous absorption permeation parameters have been derived from in vitro infinite dose studies, yet there is uncertainty in their accuracy if the applied vehicle saturates or damages the stratum corneum, or when the permeation parameters are inappropriately derived from cumulative absorption data. An approach is provided for determining penetration parameters from in vitro finite dose data. Key variables, and equations for their derivation, are identified from the literature and provide permeation parameters that use only Tmax , AUC, and AUMC from finite dose data. The equations are tested with computer-generated model data and to actual study data. Derived permeation parameters obtained from the computer model data match those used in generating the simulated finite dose data. Parameters obtained from actual study data reasonably and acceptably model the penetration profile kinetics of the study data. From in vitro finite dose absorption data, three parameters can be obtained: the diffusion transit time (td ), which characterizes the diffusion coefficient, the partition volume (Vm P), which characterizes the partition coefficient, and the permeation coefficient (Kp ). These parameters can be obtained from finite dose data without having to know the length of the diffusion pathway through the membrane. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  16. Chronological age affects the permeation of fentanyl through human skin in vitro

    DEFF Research Database (Denmark)

    Holmgaard, R; Benfeldt, E; Sorensen, J A

    2013-01-01

    AIM: To study the influence of chronological age on fentanyl permeation through human skin in vitro using static diffusion cells. Elderly individuals are known to be more sensitive to opioids and obtain higher plasma concentrations following dermal application of fentanyl compared to younger...... individuals. The influence of age - as an isolated pharmacokinetic term - on the absorption of fentanyl has not been previously studied. METHOD: Human skin from 30 female donors was mounted in static diffusion cells, and samples were collected during 48 h. Donors were divided into three age groups: ... and old age groups: 5,922 and 4,050 ng, respectively). Furthermore, the lag time and absorption rate were different between the three groups, with a significantly higher rate in the young participants versus the oldest participants. CONCLUSION: We demonstrate that fentanyl permeates the skin of young...

  17. In vitro Dermal Absorption of Hydroquinone: Protocol Validation and Applicability on Illegal Skin-Whitening Cosmetics.

    Science.gov (United States)

    Desmedt, Bart; Ates, Gamze; Courselle, Patricia; De Beer, Jacques O; Rogiers, Vera; Hendrickx, Benoit; Deconinck, Eric; De Paepe, Kristien

    2016-01-01

    In Europe, hydroquinone is a forbidden cosmetic ingredient. It is, however, still abundantly used because of its effective skin-whitening properties. The question arises as to whether the quantities of hydroquinone used become systemically available and may cause damage to human health. Dermal absorption studies can provide this information. In the EU, dermal absorption has to be assessed in vitro since the Cosmetic Regulation 1223/2009/EC forbids the use of animals. To obtain human-relevant data, a Franz diffusion cell protocol was validated using human skin. The results obtained were comparable to those from a multicentre validation study. The protocol was applied to hydroquinone and the dermal absorption ranged between 31 and 44%, which is within the range of published in vivo human values. This shows that a well-validated in vitro dermal absorption study using human skin provides relevant human data. The validated protocol was used to determine the dermal absorption of illegal skin-whitening cosmetics containing hydroquinone. All samples gave high dermal absorption values, rendering them all unsafe for human health. These results add to our knowledge of illegal cosmetics on the EU market, namely that they exhibit a negative toxicological profile and are likely to induce health problems. © 2017 S. Karger AG, Basel.

  18. Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells.

    Directory of Open Access Journals (Sweden)

    Katja Linher

    Full Text Available BACKGROUND: We have previously demonstrated that stem cells isolated from fetal porcine skin have the potential to form oocyte-like cells (OLCs in vitro. However, primordial germ cells (PGCs, which must also be specified during the stem cell differentiation to give rise to these putative oocytes at more advanced stages of culture, were not systematically characterized. The current study tested the hypothesis that a morphologically distinct population of cells derived from skin stem cells prior to OLC formation corresponds to putative PGCs, which differentiate further into more mature gametes. METHODOLOGY/PRINCIPAL FINDINGS: When induced to differentiate in an appropriate microenvironment, a subpopulation of morphologically distinct cells, some of which are alkaline phosphatase (AP-positive, also express Oct4, Fragilis, Stella, Dazl, and Vasa, which are markers indicative of germ cell formation. A known differentially methylated region (DMR within the H19 gene locus, which is demethylated in oocytes after establishment of the maternal imprint, is hypomethylated in PGC-like cells compared to undifferentiated skin-derived stem cells, suggesting that the putative germ cell population undergoes imprint erasure. Additional evidence supporting the germ cell identity of in vitro-generated PGC-like cells is that, when labeled with a Dazl-GFP reporter, these cells further differentiate into GFP-positive OLCs. SIGNIFICANCE: The ability to generate germ cell precursors from somatic stem cells may provide an in vitro model to study some of the unanswered questions surrounding early germ cell formation.

  19. Lipid nanoparticles for topical and transdermal application for alopecia treatment: development, physicochemical characterization, and in vitro release and penetration studies

    Science.gov (United States)

    Gomes, Maria João; Martins, Susana; Ferreira, Domingos; Segundo, Marcela A; Reis, Salette

    2014-01-01

    Alopecia is a dermatological disorder, commonly known as hair loss, which affects up to half of the Caucasian male population by middle age, and almost all (95%) Caucasian men by old age. Considering that alopecia affects so many people and that there is currently no scientifically proven treatment with few side effects, new drug-delivery systems able to improve alopecia therapy are urgently required. With this purpose in mind, the present study aimed to develop lipid nanoparticles (nanostructured lipid carriers) with the ability to incorporate and deliver anti-alopecia active compounds (minoxidil and finasteride) into the dermis and hair follicles. Lipid nanoparticles, prepared by ultrasonication method, showed mean particle sizes around 200 nm, which is sufficient for reaching the dermis and hair follicles, and zeta potential values around −30 mV, which indicates good physical stability. Over 28 days of storage, no significant variations in these parameters were observed, which indicates that all nanoformulations are stable in storage over that period. Cryo-scanning electron microscope measurements showed that all the lipid nanoparticles exhibited a spherical shape and a smooth surface regardless of their composition. Differential scanning calorimetry studies allowed the determination of phase transition temperatures and confirmed the recrystallization of the lipid nanoparticles (recrystallization index between 11% and 86%). A high loading efficiency was achieved for finasteride (between 70% and 90%), while less than 30% was achieved for minoxidil nanoparticles, over 28 days. Controlled release assays in physiological conditions demonstrated that nanoparticles loaded with minoxidil yielded a prolonged release, as desired. Penetration assays through pig ear skin demonstrated that nanoparticles loaded with minoxidil and finasteride had low levels of penetration. These results suggest that the proposed novel formulation presents several good characteristics

  20. Predicting skin penetration of actives from complex cosmetic formulations: an evaluation of inter formulation and inter active effects during formulation optimization for transdermal delivery.

    Science.gov (United States)

    Wiechers, J W; Watkinson, A C; Cross, S E; Roberts, M S

    2012-12-01

    Twenty products, containing a radiolabelled form of each active in typical cosmetic formulations, were made and applied to female human epidermal membranes mounted in Franz diffusion cells for 48 h under 'in use' conditions. The products consisted of combinations of five formulations (a hydro-alcoholic gel, an oil in water emulsion, a water in oil emulsion, a microemulsion and an oil) with four model drug actives (testosterone, hydrocortisone, 5-fluorouracil and ketoconazole). Steady-state flux appeared to be reached by 8 h and maintained for all products, other than for the microemulsions, consistent with the actives being present in the residual formulation on the skin at saturation. The recovery for each active at the end of the 48-h study (from a series of stratum corneum tape strips, the remaining skin, cumulative amount penetrating into the receptor solution, product washed from the skin and on the donor chamber cap) ranged from 86.5% to 100.6%. The rank order of the fluxes for the actives from the hydro-alcoholic gel is consistent with the known active molecular size and polarity determinants for maximum epidermal flux. Actives with similar steady-state (maximum) fluxes from a range of formulations had retention in the stratum corneum and similar transport rate constants through the stratum corneum. The microemulsion formulation significantly enhanced both the stratum corneum steady-state flux and transport rate constant for 5-fluorouracil, hydrocortisone and testosterone. The penetration flux of each active could be related to its size and polarity and appeared maximal when the actives in the different cosmetic formulations applied to the skin under 'in use' conditions were likely to remain in the residual product on the skin as a saturated solution after solvent evaporation. Enhanced penetration fluxes can be achieved by formulation selection and an appropriate choice/mix of emollients/adjuvants. The principles described here provide a framework for

  1. Oral administration of milk-derived phospholipids inhibits penetration of cutaneous nerve fibres into epidermis in a mouse model of acute dry skin.

    Science.gov (United States)

    Sakaguchi, A; Kamata, Y; Takahashi, N; Matsuda, H; Kosaka, R; Umehara, Y; Ogawa, H; Tominaga, M; Takamori, K

    2017-09-20

    The density of intraepidermal nerve fibres has been shown to be higher in itchy dry skin than in healthy skin, suggesting that epidermal hyperinnervation is at least partly involved in peripheral itch sensitization. We investigated whether oral administration of milk-derived phospholipids (MPLs) would inhibit epidermal hyperinnervation in a mouse model of dry skin. We found that the number of intraepidermal nerve fibres was significantly lower in the MPL group than in the control group. Expression of nerve growth factor (NGF) levels in the epidermis was significantly decreased by oral administration of MPLs, whereas expression of semaphorin (Sema)3A, a nerve repulsion factor, was increased in the MPL group. These results suggest that dietary MPLs attenuate the penetration of nerve fibres into the epidermis by reducing epidermal NGF levels and increasing Sema3A level. Thus, dietary MPLs may have beneficial effects in the prevention and/or alleviation of dry skin-induced itch by reducing intraepidermal nerve fibre density. © 2017 British Association of Dermatologists.

  2. Development of an in vivo animal model for skin penetration in hairless rats assessed by mass balance

    DEFF Research Database (Denmark)

    Simonsen, Lene; Petersen, Mads B; Benfeldt, Eva

    2002-01-01

    acid and (14)C-butyl salicylate were topically applied. Rapid and differentiated percutaneous absorption of both compounds were shown by urinary excretion data. For (14)C-salicylic acid the amount on the skin surface, in the stratum corneum and in the viable skin was determined. Total mass balance...... rat and free mobility throughout the test period. By consecutive tape stripping, monitored by measurements of transepidermal water loss and confirmed by histological examination of skin biopsies, 10 tape strippings were found to remove the stratum corneum completely. For assessment of the model, (14)C-salicylic...

  3. Suitability of skin integrity tests for dermal absorption studies in vitro.

    Science.gov (United States)

    Guth, Katharina; Schäfer-Korting, Monika; Fabian, Eric; Landsiedel, Robert; van Ravenzwaay, Ben

    2015-02-01

    Skin absorption testing in vitro is a regulatory accepted alternative method (OECD Guideline 428). Different tests can be applied to evaluate the integrity of the skin samples. Here, we compared the pre- or post-run integrity tests (transepidermal electrical resistance, TEER; transepidermal water loss, TEWL; absorption of the reference compounds water, TWF, or methylene blue, BLUE) and additionally focused on co-absorption of a (3)H-labeled internal reference standard (ISTD) as integrity parameter. The results were correlated to absorption profiles of various test compounds. Limit values of 2kΩ, 10 gm(-2)h(-1) and 4.5∗10(-3)cmh(-1) for the standard methods TEER, TEWL and TWF, respectively, allowed distinguishing between impaired and intact human skin samples in general. Single skin samples did, however, not, poorly and even inversely correlate with the test-compound absorption. In contrast, results with ISTD (e.g. (3)H-testosterone) were highly correlated to the absorption of (14)C-labeled test compounds. Importantly, ISTD did not influence analytics or absorption of test compounds. Therefore, ISTD, especially when adjusted to the physico-chemical properties of test compounds, is a promising concept to assess the integrity of skin samples during the whole course of absorption experiments. However, a historical control dataset is yet necessary for a potential routine application. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  4. Keratinocyte cultures from involved skin in vitiligo patients show an impaired in vitro behaviour.

    Science.gov (United States)

    Bondanza, Sergio; Maurelli, Riccardo; Paterna, Patrizia; Migliore, Eleonora; Giacomo, Fabio Di; Primavera, Giovanni; Paionni, Emanuel; Dellambra, Elena; Guerra, Liliana

    2007-08-01

    Vitiligo depigmentation is considered a consequence of either melanocyte disappearance or loss of functioning melanocytes in the involved areas. However, it has been reported that keratinocytes in involved vitiligo skin are damaged too. Based on this evidence, we evaluated the in vitro behaviour, in life span cultures, of involved and uninvolved vitiligo keratinocytes and their expression of proliferation, differentiation and senescence markers. An additional purpose was to investigate whether vitiligo keratinocytes from depigmented skin are able to sustain survival and growth of normal melanocytes (when added in co-culture experiments), as normal human keratinocytes manage to do. Our results demonstrate that almost all involved vitiligo keratinocytes have a shorter life span in vitro than the uninvolved cells and all of them do not maintain melanocytes in culture in a physiological ratio. Modification of proliferation and senescence marker expression also occurs. Indeed, we detected low initial expression levels of the senescence marker p16 in involved vitiligo keratinocytes, despite their shorter in vitro life span, and increased expression of proliferating cell nuclear antigen and p53. This preliminary analysis of a small number of in vitro cultured vitiligo keratinocytes suggests an impaired senescence process in lesional vitiligo keratinocytes and attempts to regulate it.

  5. The effect of local hyperglycemia on skin cells in vitro and on wound healing in euglycemic rats

    DEFF Research Database (Denmark)

    Kruse, Carla R; Singh, Mansher; Sørensen, Jens A

    2016-01-01

    BACKGROUND: Multiple previous studies have established that high systemic blood glucose concentration impairs skin wound healing. However, the effects of local hyperglycemia on wound healing are not well defined. Comprehensive animal studies and in vitro studies using both fibroblasts and keratin......BACKGROUND: Multiple previous studies have established that high systemic blood glucose concentration impairs skin wound healing. However, the effects of local hyperglycemia on wound healing are not well defined. Comprehensive animal studies and in vitro studies using both fibroblasts...

  6. In vitro study of RRS HA injectable mesotherapy/biorevitalization product on human skin fibroblasts and its clinical utilization.

    Science.gov (United States)

    Deglesne, Pierre-Antoine; Arroyo, Rodrigo; Ranneva, Evgeniya; Deprez, Philippe

    2016-01-01

    Mesotherapy/biorevitalization with hyaluronic acid (HA) is a treatment approach currently used for skin rejuvenation. Various products with a wide range of polycomponent formulations are available on the market. Most of these formulations contain noncross-linked HA in combination with a biorevitalization cocktail, formed by various amounts of vitamins, minerals, amino acids, nucleotides, coenzymes, and antioxidants. Although ingredients are very similar among the different products, in vitro and clinical effects may vary substantially. There is a real need for better characterization of these products in terms of their action on human skin or in vitro skin models. In this study, we analyzed the effect of the RRS(®) (Repairs, Refills, Stimulates) HA injectable medical device on human skin fibroblasts in vitro. Skin fibroblast viability and its capacity to induce the production of key extracellular matrix were evaluated in the presence of different concentrations of RRS HA injectable. Viability was evaluated through colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay, and key extracellular matrix genes, type I collagen and elastin, were quantified by quantitative polymerase chain reaction. Results demonstrated that RRS HA injectable could promote human skin fibroblast viability (+15%) and increase fibroblast gene expression of type I collagen and elastin by 9.7-fold and 14-fold in vitro, respectively. These results demonstrate that mesotherapy/biorevitalization products can, at least in vitro, effectively modulate human skin fibroblasts.

  7. Release and in vitro skin permeation of polyphenols from cosmetic emulsions.

    Science.gov (United States)

    Zillich, O V; Schweiggert-Weisz, U; Hasenkopf, K; Eisner, P; Kerscher, M

    2013-10-01

    Polyphenols are natural antioxidants, which can inhibit oxidative chain reactions in human skin and prevent therefore some skin diseases and premature ageing. A prerequisite of this behaviour is their permeation through the skin barrier, in particular the stratum corneum (SC). In this study, we investigated the skin permeation kinetic of polyphenols, incorporated to semisolid emulsions, and the release of polyphenols from the emulsions. Mixtures of model substances, consisting of catechin, epigallocatechin gallate (EGCG), resveratrol, quercetin, rutin and protocatechuic acid (PCA), were formulated into o/w emulsions with different oil phase content. The in vitro experiments were carried out in Franz-type diffusion cells by means of ex vivo pig skin and a cellulose membrane. The increased oil content in the emulsion led to a significant decrease in initial release coefficients (K(r)), diffusion coefficients within the formulation (D(v)) and skin permeation coefficients (K(p)), respectively. The study considered the dependence of K(r) on molecular weight and lipophilicity of polyphenolics. For both more hydrophilic and more lipophilic substance groups, the values for K(r) were inverse proportional to molecular weight. For catechin, quercetin, rutin, resveratrol and PCA, a good correlation between K(p) and K(r) parameters was obtained. The most permeable substance was PCA (K(p) = 1.2·10(-3) cm h(-1)), and the least permeable was quercetin (K(p) = 1.5·10(-5) cm h(-1)). All substances could pass the SC barrier and were found mostly in the epidermis and dermis, confirming the potential of polyphenols as anti-ageing active cosmetic ingredients. © 2013 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  8. Characterization and In Vitro Skin Permeation of Meloxicam-Loaded Liposomes versus Transfersomes

    Directory of Open Access Journals (Sweden)

    Sureewan Duangjit

    2011-01-01

    Full Text Available The goal of this study was to develop and evaluate the potential use of liposome and transfersome vesicles in the transdermal drug delivery of meloxicam (MX. MX-loaded vesicles were prepared and evaluated for particle size, zeta potential, entrapment efficiency (%EE, loading efficiency, stability, and in vitro skin permeation. The vesicles were spherical in structure, 90 to 140 nm in size, and negatively charged (−23 to −43 mV. The %EE of MX in the vesicles ranged from 40 to 70%. Transfersomes provided a significantly higher skin permeation of MX compared to liposomes. Fourier Transform Infrared Spectroscopy (FT-IR and Differential Scanning Calorimetry (DSC analysis indicated that the application of transfersomes significantly disrupted the stratum corneum lipid. Our research suggests that MX-loaded transfersomes can be potentially used as a transdermal drug delivery system.

  9. Skin fibroblasts from individuals hemizygous for the familial adenopolyposis susceptibility gene show delayed crisis in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Chen, S.; Kazim, D.; Kraveka, J.; Pollack, R.E. (Columbia Univ., New York, NY (USA))

    1989-03-01

    Normal human fibroblast cells have not been reported to escape crisis--that is they die after about 24 doublings in culture. The authors have been studying the growth properties of skin fibroblast cells from persons in families with familial adenopolyposis of the colon (FAP). An individual hemizygous at the FAP locus will develop hyperplasia of the colonic epithelium followed by colonic polyps, both at an early age. Polyps themselves still retain a single functional FAP allele. A mutation or deletion in this allele in a polyp is hypothesized to lead to further loss of growth control; thus, a tumor is formed. They found that the in vitro life-span of skin fibroblast cells from FAP individuals and from some asymptomatic children were markedly extended when compared with normal individuals.

  10. Candesartan cilexetil microemulsions for transdermal delivery: formulation, in-vitro skin permeation and stability assessment.

    Science.gov (United States)

    Malakar, Jadupati; Basu, Aalok; Nayak, Amit Kumar

    2014-01-01

    The work investigates the formulation and evaluation of microemulsions containing olive oil, Tween 80 and isopropyl alcohol for transdermal candesartan cilexetil delivery. The pseudoternary phase diagram was constructed to determine composition of microemulsions. These formulated microemulsions were evaluated for in vitro skin permeation and stability. The microemulsion containing 72 % olive oil, 8 % water, 15 % Tween 80, and 5 % isopropylalcohol showed maximum viscosity of 29.54±0.32 mPas, average small droplet size of 180.90 nm, smaller polydispersity index of 0.37, zeta potential of -12.20 and maximum candesartan cilexetil permeation flux of 0.49±0.05 μg/cm2/h through excised porcine skin. The degradation of candesartan cilexetil microemulsions after 3 months storage was found low and its shelf-life was calculated as 3.92 years at room temperature.

  11. In Vitro and In Vivo Enhancement of Antitumoral Activity of Liposomal Antisense Oligonucleotides by Cineole as a Chemical Penetration Enhancer

    Directory of Open Access Journals (Sweden)

    Hamid Reza Moghimi

    2015-01-01

    Full Text Available Cellular uptake and cytoplasmic release of liposomal antisense oligonucleotides (AsODNs, which can act as rate-limiting steps, are still remained to be completely optimized. Here, the possibility of enhancing such processes at cellular and animal levels by cineole, as a penetration enhancer, was investigated. A cationic nanoliposome containing an AsODN against PKC-α and a cineole-containing nanoliposome were prepared and characterized. The effect of nanoliposomal cineole on sequence-specific cytotoxicity of nanoliposomal AsODN against A549, was studied in vitro (MTT, flow cytometry, fluorescence microscopy, and real time PCR and in vivo (xenograft lung tumor in nude mice using different concentrations and treatment times. Results showed specific cytotoxicity of nanoliposomal AsODN was increased significantly from 11% to 25% when A549 cells were exposed to 10 µg/mL cineole for 1 or 4 hours. This inhibitory effect was further increased to about 40% when the concentration was increased to 40 µg/mL for 1 hour. In animal studies, cineole significantly decreased the tumor volume (about 75% and increased its doubling time from 13 days to 31 days. A linear relationship exists between cineole concentration and its enhancement effects. Finally it was concluded that cineole, and possibly other membrane fluidizers, can improve nanoliposomal gene therapy at cellular and animal levels.

  12. The in vitro antimicrobial activity of wound and skin cleansers at nontoxic concentrations.

    Science.gov (United States)

    Rani, Suriani Abdul; Hoon, Russell; Najafi, Ramin Ron; Khosrovi, Behzad; Wang, Lu; Debabov, Dmitri

    2014-02-01

    To determine in vitro antibacterial activity of commercially available skin, wound, and skin/wound cleansers at cell-safe (nontoxic) concentrations. Saline and 19 other commercial wound and skin cleansers were evaluated for cytotoxic effects on mouse dermal fibroblasts. Cells were exposed to serial 10-fold dilutions of each cleanser until treatment-induced cytotoxicity was comparable to the baseline cytotoxicity of unexposed control fibroblasts. Antimicrobial "time-kill" kinetics of these test concentrations of cleansers was tested against methicillin-resistant Staphylococcus aureus. The experimental design allowed calculation of relative cytotoxicity indexes ranging from 0 to 100,000. Shur-Clens Restore Wound Cleanser (ConvaTec, Skillman, New Jersey) and saline were found to be the least toxic (toxicity index: 0); Hibiclens (Mölnlycke Health Care, Norcross, Georgia), Restore Skin Cleanser (Hollister Inc, Libertyville, Illinois), and Betadine Surgical Scrub (Pursue Products LP, Stamford, Connecticut) were found to be the most toxic (toxicity index: 10,000). At noncytotoxic concentrations, NeutroPhase (NovaBay Pharmaceuticals Inc, Emeryville, California) was the most rapidly bactericidal, achieving a 4-log reduction in colony-forming units in less than 1 minute. Puracyn (Innovacyn Inc, Rialto, California) was next at 30 minutes, whereas most of the agents tested required more than 24 hours. Wound healing depends on controlling bacterial balance while maintaining the viability of the healing tissues. In vitro toxicity indexes provide helpful guidelines subsequent to in vivo evaluations and clinical applications. The study findings suggest that NeutroPhase, in contrast with many commercially available wound cleansers, has rapid bactericidal activity at concentrations that are safe for human cells.

  13. Simple Amides of Oleanolic Acid as Effective Penetration Enhancers

    Science.gov (United States)

    Bednarczyk-Cwynar, Barbara; Partyka, Danuta; Zaprutko, Lucjusz

    2015-01-01

    Transdermal transport is now becoming one of the most convenient and safe pathways for drug delivery. In some cases it is necessary to use skin penetration enhancers in order to allow for the transdermal transport of drugs that are otherwise insufficiently skin-permeable. A series of oleanolic acid amides as potential transdermal penetration enhancers was formed by multistep synthesis and the synthesis of all newly prepared compounds is presented. The synthetized amides of oleanolic acid were tested for their in vitro penetration promoter activity. The above activity was evaluated by means of using the Fürst method. The relationships between the chemical structure of the studied compounds and penetration activity are presented. PMID:26010090

  14. Bioactive reagents used in mesotherapy for skin rejuvenation in vivo induce diverse physiological processes in human skin fibroblasts in vitro- a pilot study.

    Science.gov (United States)

    Jäger, Claudia; Brenner, Christiane; Habicht, Jüri; Wallich, Reinhard

    2012-01-01

    The promise of mesotherapy is maintenance and/or recovery of a youthful skin with a firm, bright and moisturized texture. Currently applied medications employ microinjections of hyaluronic acid, vitamins, minerals and amino acids into the superficial layer of the skin. However, the molecular and cellular processes underlying mesotherapy are still elusive. Here we analysed the effect of five distinct medication formulas on pivotal parameters involved in skin ageing, that is collagen expression, cell proliferation and morphological changes using normal human skin fibroblast cultures in vitro. Whereas in the presence of hyaluronic acid, NCTF135(®) and NCTF135HA(®) , cell proliferation was comparable to control cultures; however, with higher expression of collagen type-1, matrix metalloproteinase-1 and tissue inhibitor of matrix metalloproteinase-1, addition of Soluvit(®) N and Meso-BK led to apoptosis and/or necrosis of human fibroblasts. The data indicate that bioactive reagents currently applied for skin rejuvenation elicit strikingly divergent physiological processes in human skin fibroblast in vitro. © 2011 John Wiley & Sons A/S.

  15. Identification and partial sequencing of a crocodile poxvirus associated with deeply penetrating skin lesions in farmed Nile crocodiles, Crocodylus niloticus

    Directory of Open Access Journals (Sweden)

    F.W. Huchzermeyer

    2009-09-01

    Full Text Available When large numbers of crocodile skins were downgraded because of the presence of small pin pricklike holes, collapsed epidermal cysts were found deep in the dermis of juvenile crocodiles while forming cysts were observed in hatchlings. Histopathology of these forming cysts showed the presence of intracytoplasmic inclusions in proliferating and ballooning epidermal cells. Pox virions were seen in electron microscope preparations made from the scabs of such early lesions. The partial sequencing of virus material from scrapings of these lesions and comparison of it with the published sequence of crocodile poxvirus showed the virus associated with the deep lesions to be closely related, but different. To differentiate between the two forms of crocodile pox infection it is suggested that the previously known form should be called ''classical crocodile pox'' and the newly discovered form ''atypical crocodile pox''. The application of strict hygiene measures brought about a decline in the percentage of downgraded skins.

  16. Identification and partial sequencing of a crocodile poxvirus associated with deeply penetrating skin lesions in farmed Nile crocodiles, Crocodylus niloticus.

    Science.gov (United States)

    Huchzermeyer, F W; Wallace, D B; Putterill, J F; Gerdes, G H

    2009-09-01

    When large numbers of crocodile skins were downgraded because of the presence of small pin prick-like holes, collapsed epidermal cysts were found deep in the dermis of juvenile crocodiles while forming cysts were observed in hatchlings. Histopathology of these forming cysts showed the presence of intracytoplasmic inclusions in proliferating and ballooning epidermal cells. Pox virions were seen in electron microscope preparations made from the scabs of such early lesions. The partial sequencing of virus material from scrapings of these lesions and comparison of it with the published sequence of crocodile poxvirus showed the virus associated with the deep lesions to be closely related, but different. To differentiate between the two forms of crocodile pox infection it is suggested that the previously known form should be called "classical crocodile pox" and the newly discovered form "atypical crocodile pox". The application of strict hygiene measures brought about a decline in the percentage of downgraded skins.

  17. In vitro incidence of fibular penetration with and without the use of a jig during tibial plateau leveling osteotomy.

    Science.gov (United States)

    Flynn, Patrick; Duncan, Colleen G; Palmer, Ross H; Duerr, Felix M

    2014-05-01

    To determine the incidence of fibular penetration during placement of the Synthes® locking TPLO plate with and without the use of a jig. Cadaveric, experimental study. Cadaveric paired pelvic limbs (n = 8) from skeletally mature dogs. Limbs were assigned to 1 of 2 groups (jig-less-TPLO = no jig used; jig-TPLO = jig used). Synthes® locking TPLO plates were applied using proximal screws of excessive length to facilitate identification of fibular penetration. Screw tip locations were identified by dissection and frequency of fibular penetration was compared between groups. None of the jig-TPLO limbs and 6 (75%) of jig-less-TPLO limbs had fibular penetration, a difference that was statistically significant. Fibular penetration was most frequently associated with the most proximal screw. A significantly higher incidence of fibular penetration occurs when Synthes® locking TPLO is performed without use of a jig. © Copyright 2014 by The American College of Veterinary Surgeons.

  18. Microleakage and penetration depth of three types of materials in fissure sealant: self-etching primer vs etching: an in vitro study.

    Science.gov (United States)

    Gillet, D; Nancy, J; Dupuis, V; Dorignac, G

    2002-01-01

    Clinical preventive procedures must be done after a risk assessment. One of the risk factors is the occlusal morphology of the posterior teeth. These caries-free fissures must be sealed. This first in vitro experimentation of the study evaluated the microleakage and the penetration depth of three types of materials by Vivadent: Helioseal F, Tetric, Tetric Flow. The teeth were etched with phosphoric acid and bonded using a one bottle bonding in order to determine the best material for the sealing of the fissure. The depth of penetration of fuschine dye as well as that of the tested material was measured with a grid. The results, compared to the depth of the fissures, are expressed in percentage of penetration. The results were as follows: penetration of fuschine dye: 0% for the 2 composites, 100% for Helioseal F; penetration of the materials: 96.90% for Helioseal F, 70.82 for Tetric and 86.10 for Tetric Flow (significant difference, Wilcoxon test = 0.0105). In this first in vitro study, Tetric Flow shows no microleakage and is more efficient when compared to Helioseal F and Tetric in obturating deep fissures of non carious bicuspids. The second experiment of the study evaluated the microleakage and the penetration depth of Tetric Flow when it is bonded by two different methods: Group 1: total etch (phosphoric acid) and Scotch-bond 1 (3M), and Group 2: self-etching primer with Prompt (Espe). There was no significant difference (p > 0.03) between classical bonding vs self-etching primer. The self-etching primer Prompt is very efficient vs phosphoric acid in obturating the fissures of non carious bicuspids with Tetric Flow. It is concluded that for prevention by sealing, using a flowable ceromer (Tetric Flow) with the self-etching (Prompt), is a really good technique.

  19. An in vitro model for detecting skin irritants: methyl green-pyronine staining of human skin explant cultures

    NARCIS (Netherlands)

    Jacobs, J. J. L.; Lehé, C.; Cammans, K. D. A.; Das, P. K.; Elliott, G. R.

    2002-01-01

    We evaluated the potential of human organotypic skin explant cultures (hOSECs) for screening skin irritants. Test chemicals were applied to the epidermis of the skin explants which were incubated for 4, 24 or 48 h in tissue culture medium. A decrease in epidermal RNA staining, visualised in frozen

  20. Differential susceptibility of primary cultured human skin cells to hypericin PDT in an in vitro model.

    Science.gov (United States)

    Popovic, A; Wiggins, T; Davids, L M

    2015-08-01

    Skin cancer is the most common cancer worldwide, and its incidence rate in South Africa is increasing. Photodynamic therapy (PDT) has been shown to be an effective treatment modality, through topical administration, for treatment of non-melanoma skin cancers. Our group investigates hypericin-induced PDT (HYP-PDT) for the treatment of both non-melanoma and melanoma skin cancers. However, a prerequisite for effective cancer treatments is efficient and selective targeting of the tumoral cells with minimal collateral damage to the surrounding normal cells, as it is well established that cancer therapies have bystander effects on normal cells in the body, often causing undesirable side effects. The aim of this study was to investigate the cellular and molecular effects of HYP-PDT on normal primary human keratinocytes (Kc), melanocytes (Mc) and fibroblasts (Fb) in an in vitro tissue culture model which represented both the epidermal and dermal cellular compartments of human skin. Cell viability analysis revealed a differential cytotoxic response to a range of HYP-PDT doses in all the human skin cell types, showing that Fb (LD50=1.75μM) were the most susceptible to HYP-PDT, followed by Mc (LD50=3.5μM) and Kc (LD50>4μM HYP-PDT) These results correlated with the morphological analysis which displayed distinct morphological changes in Fb and Mc, 24h post treatment with non-lethal (1μM) and lethal (3μM) doses of HYP-PDT, but the highest HYP-PDT doses had no effect on Kc morphology. Fluorescent microscopy displayed cytoplasmic localization of HYP in all the 3 skin cell types and additionally, HYP was excluded from the nuclei in all the cell types. Intracellular ROS levels measured in Fb at 3μM HYP-PDT, displayed a significant 3.8 fold (pPDT. These results depict a differential response to HYP-PDT by different human skin cells thus highlighting the efficacy and indeed, the potential bystander effect of if administered in vivo. This study contributes toward our knowledge

  1. Assessment of penetrant and vehicle mixture properties on transdermal permeability using a mixed effect pharmacokinetic model of ex vivo porcine skin.

    Science.gov (United States)

    Chittenden, Jason T; Riviere, Jim E

    2016-10-01

    The accurate prediction of the rate and extent of transdermal absorption from topical exposure to chemical mixtures would be beneficial in risk assessment and drug delivery applications. The isolated perfused porcine skin flap (IPPSF) has been used as an ex vivo model for assessing transdermal absorption from topical exposures. A mixed effect, pharmacokinetic tissue model was used to model finite dose, transdermal, absorption data from IPPSF experiments for 12 penetrants dosed in up to 10 different vehicles. The model was able to identify permeability constant, while accounting for between and within unit variability, across the entire data set. This approach provides a platform for exploring the relationship between covariates (chemical descriptors and functions thereof) and the model parameters. Successive models were employed that reduced the overall variability in the parameter estimate by modeling the parameters as functions of the covariates. Log kp was initially modeled as a function of LogP and MW of the pure penetrant (adjusted r2  = 0.48). The addition of mixture factors to account for the different dosing vehicles further improved the relationship: to r2  = 0.56 with Connolly molecular area (CMA) and r2  = 0.78 with the further addition of total polar surface area difference (TPSAd). The pharmacokinetic model and quantitative structure property relationship (QSPR) developed for the IPPSF may be relevant to clinical transdermal formulation development. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  2. In vitro study of RRS HA injectable mesotherapy/biorevitalization product on human skin fibroblasts and its clinical utilization

    Directory of Open Access Journals (Sweden)

    Deglesne PA

    2016-02-01

    Full Text Available Pierre-Antoine Deglesne,* Rodrigo Arroyo,* Evgeniya Ranneva, Philippe Deprez Research and Development, SKIN TECH PHARMA GROUP, Castelló d'Empúries, Spain  *These authors contributed equally to this work Abstract: Mesotherapy/biorevitalization with hyaluronic acid (HA is a treatment approach currently used for skin rejuvenation. Various products with a wide range of polycomponent formulations are available on the market. Most of these formulations contain noncross-linked HA in combination with a biorevitalization cocktail, formed by various amounts of vitamins, minerals, amino acids, nucleotides, coenzymes, and antioxidants. Although ingredients are very similar among the different products, in vitro and clinical effects may vary substantially. There is a real need for better characterization of these products in terms of their action on human skin or in vitro skin models. In this study, we analyzed the effect of the RRS® (Repairs, Refills, Stimulates HA injectable medical device on human skin fibroblasts in vitro. Skin fibroblast viability and its capacity to induce the production of key extracellular matrix were evaluated in the presence of different concentrations of RRS HA injectable. Viability was evaluated through colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, and key extracellular matrix genes, type I collagen and elastin, were quantified by quantitative polymerase chain reaction. Results demonstrated that RRS HA injectable could promote human skin fibroblast viability (+15% and increase fibroblast gene expression of type I collagen and elastin by 9.7-fold and 14-fold in vitro, respectively. These results demonstrate that mesotherapy/biorevitalization products can, at least in vitro, effectively modulate human skin fibroblasts.Keywords: mesotherapy, medical device, RRS, collagen, elastin, extracellular matrix

  3. Evaluation of light-emitting diodes (LED) effect on skin biology (in vitro study).

    Science.gov (United States)

    Chabert, R; Fouque, L; Pinacolo, S; Garcia-Gimenez, N; Bonnans, M; Cucumel, K; Domloge, N

    2015-11-01

    Interest in anti-aging approaches has grown significantly in recent years. The most popular are the non invasive methods to decrease the signs of aging. One such method is LED-based therapy. This study investigated the potential of two different wavelengths, 590 nm and 630 nm, combined or not, in the photobiomodulation of proteins involved in the slowdown of the skin aging. These in vitro results on cell viability, cell shape, and mitochondrial function support and build on previous studies suggested that LED treatment is safe. Regarding its biological functions, our data indicated that the combination of two different wavelengths acted in synergy to enhance the impact of each irradiation alone. Combined, the LED wavelengths could improve in vitro the cell shape, the cell proliferation, and the level of major proteins involved in the healing process. These benefits may lead to reinforcement of the skin organization and structure. This hypothesis will be checked in future clinical studies. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  4. Zingiber cassumunar blended patches for skin application: Formulation, physicochemical properties, and in vitro studies

    Directory of Open Access Journals (Sweden)

    Jirapornchai Suksaeree

    2015-07-01

    Full Text Available Our work was to study the preparation, physicochemical characterization, and in vitro characteristic of Zingiber cassumunar blended patches. The Z. cassumunar blended patches incorporating Z. cassumunar Roxb. also known as Plai were prepared from chitosan and polyvinyl alcohol with glycerin as plasticizer. They were prepared by adding all ingredients in a beaker and homogeneously mixing them. Then, they were transferred into Petri-dish and dried in hot air oven. The hydrophilic nature of the Z. cassumunar blended patches was confirmed by the moisture uptake, swelling ratio, erosion, and porosity values. The FTIR, DSC, XRD, and SEM studies showed revealed blended patches with amorphous region that was homogeneously smooth and compact in both surface and cross section dimensions. They exhibited controlled the release behavior of (E-4-(3′,4′-dimethoxyphenyl but-3-en-l-ol (compound D that is the main active compound in Z. cassumunar for anti-inflammation activity. However, in in vitro skin permeation study, the compound D was accumulated in newborn pig skin more than in the receptor medium. Thus, the blended patches showed the suitable entrapment and controlled release of compound D. Accordingly, we have demonstrated that such chitosan and polyvinyl alcohol formulated patches might be developed for medical use.

  5. Alfuzosin hydrochloride transdermal films: evaluation of physicochemical, in vitro human cadaver skin permeation and thermodynamic parameters

    Directory of Open Access Journals (Sweden)

    Satyanarayan Pattnaik

    2009-12-01

    Full Text Available Purpose: The main objective of the investigation was to develop a transdermal therapeutic system for alfuzosin hydrochloride and to study the effects of polymeric system and loading dose on the in vitro skin permeation pattern. Materials and methods: Principles of experimental design have been exploited to develop the dosage form. Ratio of ethyl cellulose (EC and polyvinyl pyrrolidone (PVP and loading dose were selected as independent variables and their influence on the cumulative amount of alfuzosin hydrochloride permeated per cm2 of human cadaver skin at 24 h (Q24, permeation flux (J and steady state permeability coefficient (P SS were studied using experimental design. Various physicochemical parameters of the transdermal films were also evaluated. Activation energy for in vitro transdermal permeation has been estimated. Results: Ratio of EC and PVP was found to be the main influential factor for all the dependent variables studied. Drug loading dose was also found to influence the dependent variables but to a lesser extent. Physicochemical parameters of the prepared films were evaluated and found satisfactory. Activation energy for alfuzosin permeation has also been estimated and reported. Conclusion: The therapeutic system was found to be dermatologically non-irritant and hence, a therapeutically effective amount of alfuzosin hydrochloride can be delivered via a transdermal route.

  6. In vitro bioartificial skin culture model of tissue rejection and inflammatory/immune mechanisms.

    Science.gov (United States)

    Strande, L F; Foley, S T; Doolin, E J; Hewitt, C W

    1997-06-01

    We hypothesized that an in vitro bioartificial skin rejection model using living LSEs grown in tissue culture could be developed for the study of autologous, allogenic, and/or xenogeneic inflammatory/immune mechanisms and topical immunosuppressive drugs. Human fibroblasts were mixed with type 1 rat-tail collagen to form a matrix (4 to 5 days), on which human keratinocytes were seeded. After a keratinocyte monolayer formed, CT cultures were raised to the air-liquid interface for continued growth. In the REJ LSE model, immunocytes isolated from human blood were seeded on top of the NHEK monolayer at the time of air-lifting. Thickness measurements of the acellular keratin and keratinocyte layers, and nuclear/cytoplasmic ratios, in both CT and REJ were made using digital image analysis. Immunostaining with anticytokeratin demonstrated a viable, keratin-producing epidermal layer; staining with anti-TGF-beta suggested a role for this cytokine in the rejection or wound-healing process. The LSE appeared histologically similar to normal human epidermis. Immunocytes added to the REJ cultures caused an obvious rejection response and were clearly identifiable in the gels as CD45+ staining cells. The LSE model appears promising for the study of immune/inflammatory mechanisms, thermal injury, screening antirejection agents that might be applied topically and as an in vitro replacement for skin graft studies in animals.

  7. In Vitro Differentiation Potential of Human Placenta Derived Cells into Skin Cells

    Directory of Open Access Journals (Sweden)

    Ruhma Mahmood

    2015-01-01

    Full Text Available Skin autografting is the most viable and aesthetic technique for treatment of extensive burns; however, this practice has potential limitations. Harvesting cells from neonatal sources (such as placental tissue is a simple, inexpensive, and noninvasive procedure. In the current study authors sought to evaluate in vitro potential of human placenta derived stem cells to develop into skin-like cells. After extensive washing, amniotic membrane and umbilical cord tissue were separated to harvest amniotic epithelial cells (AECs and umbilical cord mesenchymal stem cells (UC-MSCs, respectively. Both types of cells were characterized for the expression of embryonic lineage markers and their growth characteristics were determined. AECs and UC-MSCs were induced to differentiate into keratinocytes-like and dermal fibroblasts-like cells, respectively. After induction, morphological changes were detected by microscopy. The differentiation potential was further assessed using immunostaining and RT-PCR analyses. AECs were positive for cytokeratins and E-Cadherin while UC-MSCs were positive for fibroblast specific makers. AECs differentiated into keratinocytes-like cells showed positive expression of keratinocyte specific cytokeratins, involucrin, and loricrin. UC-MSCs differentiated into dermal fibroblast-like cells indicated expression of collagen type 3, desmin, FGF-7, fibroblast activation protein alpha, procollagen-1, and vimentin. In conclusion, placenta is a potential source of cells to develop into skin-like cells.

  8. Evidence from the very beginning: endoglandular trophoblasts penetrate and replace uterine glands in situ and in vitro.

    Science.gov (United States)

    Moser, G; Weiss, G; Gauster, M; Sundl, M; Huppertz, B

    2015-12-01

    How is histiotrophic nutrition of the embryo secured during the first trimester of pregnancy? Rather than specifically focusing on invasion into spiral arteries, extravillous trophoblasts also invade into uterine glands (endoglandular trophoblast) from the very beginning and open them toward the intervillous space. Extravillous trophoblasts can be found in close contact and within the lumen of uterine glands, sometimes replacing glandular epithelial cells. As well as extensive screening of specimens from first trimester placentation sites in situ we used a previously established three-dimensional co-culture in vitro model system of first trimester villous explants with non-invaded decidua parietalis. First trimester placentas were obtained from elective terminations of pregnancies (n = 48) at 5-11 weeks of gestational age. A subset was processed for confrontation co-culture (n = 31). Invaded decidua basalis was obtained from 20 placentas. All tissues were sectioned, subsequently immunostained and immunodoublestained with antibodies against keratin 7 (KRT7), major histocompatibility complex, class I, G (HLA-G), matrix metallopeptidase 9 (MMP9), von Willebrand factor (VWF) and the appropriate Immunoglobulin G (IgG) negative controls. Replacement of endothelial/epithelial cells by extravillous trophoblasts was quantified semi-quantitatively. Additionally, hematoxylin and eosin-stained archival specimens from early implantation sites were assessed. The earliest available specimen was from around Day 10 after conception; already at this stage trophoblasts had penetrated into uterine glands and had started to replace the epithelium of the glands. Endoglandular trophoblasts replaced uterine glands in vitro and in situ and could be found in the lumen of invaded glands. Quantitative analysis revealed significantly more replacement of epithelial cells in glands (63.8 ± 22.1%) compared with endothelial cells in vessels (26.4 ± 8.8%). Accumulated detached glandular

  9. Ultrasound-mediated transdermal drug delivery of fluorescent nanoparticles and hyaluronic acid into porcine skin in vitro

    Science.gov (United States)

    Wang, Huan-Lei; Fan, Peng-Fei; Guo, Xia-Sheng; Tu, Juan; Ma, Yong; Zhang, Dong

    2016-12-01

    Transdermal drug delivery (TDD) can effectively bypass the first-pass effect. In this paper, ultrasound-facilitated TDD on fresh porcine skin was studied under various acoustic parameters, including frequency, amplitude, and exposure time. The delivery of yellow-green fluorescent nanoparticles and high molecular weight hyaluronic acid (HA) in the skin samples was observed by laser confocal microscopy and ultraviolet spectrometry, respectively. The results showed that, with the application of ultrasound exposures, the permeability of the skin to these markers (e.g., their penetration depth and concentration) could be raised above its passive diffusion permeability. Moreover, ultrasound-facilitated TDD was also tested with/without the presence of ultrasound contrast agents (UCAs). When the ultrasound was applied without UCAs, low ultrasound frequency will give a better drug delivery effect than high frequency, but the penetration depth was less likely to exceed 200 μm. However, with the help of the ultrasound-induced microbubble cavitation effect, both the penetration depth and concentration in the skin were significantly enhanced even more. The best ultrasound-facilitated TDD could be achieved with a drug penetration depth of over 600 μm, and the penetration concentrations of fluorescent nanoparticles and HA increased up to about 4-5 folds. In order to get better understanding of ultrasound-facilitated TDD, scanning electron microscopy was used to examine the surface morphology of skin samples, which showed that the skin structure changed greatly under the treatment of ultrasound and UCA. The present work suggests that, for TDD applications (e.g., nanoparticle drug carriers, transdermal patches and cosmetics), protocols and methods presented in this paper are potentially useful. Project partially supported by the National Natural Science Foundation of China (Grant Nos. 81127901, 81227004, 81473692, 81673995, 11374155, 11574156, 11274170, 11274176, 11474001

  10. Use of Curcuma longa in cosmetics: extraction of curcuminoid pigments, development of formulations, and in vitro skin permeation studies

    Directory of Open Access Journals (Sweden)

    Gisele Mara Silva Gonçalves

    2014-12-01

    Full Text Available Curcuma longais a ginger family aromatic herb (Zingiberaceae whose rhizomes contain curcuminoid pigments, including curcumin, a compound known for its anti-inflammatory effects. The objective of this study was to obtain curcuminoid-rich extracts, develop topical formulations thereof, and assess the stability and skin permeation of these formulations. Curcuma longa extracts were obtained and used to develop formulations. Skin permeation studies were conducted in a modified Franz diffusion cell system, and skin retention of curcuminoid pigments was quantified in pig ear membrane. Prepared urea-containing gel-cream formulations were unstable, whereas all others had satisfactory stability and pseudoplastic rheological behavior. The amount of curcuminoid pigments recovered from the receptor solution was negligible. The skin concentration of curcuminoid pigments retained was positive (>20 µg/g of skin, mostly in the stratum corneum, considering the low skin permeability of curcumin. We conclude that development of topical formulations containing curcumin or Curcuma longaextract is feasible, as long as adjuvants are added to improve preservation and durability. The formulations developed in this study enabled penetration of curcumin limited to the superficial layers of the skin and then possibly without a risk of systemic action, thus permitting local use as a topical anti-inflammatory.

  11. Resveratrol-Loaded Lipid Nanocarriers: Correlation between In Vitro Occlusion Factor and In Vivo Skin Hydrating Effect

    Directory of Open Access Journals (Sweden)

    Lucia Montenegro

    2017-12-01

    Full Text Available Lipid nanocarriers show occlusive properties that may be related to their ability to improve skin hydration. The aim of this work was to evaluate the relationship between in vitro occlusion factor and in vivo skin hydration for three types of lipid nanocarriers: nanoemulsions (NEs, solid lipid nanoparticles (SLNs and nanostructured lipid carriers (NLCs. These lipid nanocarriers were loaded with trans-resveratrol (RSV and incorporated in gel vehicles. In vitro occlusion factor was in the order SLNs > NLCs > NEs. Gels containing unloaded or RSV loaded lipid nanocarriers were applied on the back of a hand of 12 healthy volunteers twice a day for one week, recording skin hydration changes using the instrument Soft Plus. An increase of skin hydration was observed for all lipid nanocarriers (SLNs > NLCs > NEs. RSV loading into these nanocarriers did not affect in vitro and in vivo lipid nanocarriers effects. A linear relationship (r2 = 0.969 was observed between occlusion factor and in vivo increase of skin hydration. Therefore, the results of this study showed the feasibility of using the occlusion factor to predict in vivo skin hydration resulting from topical application of different lipid nanocarriers loading an active ingredient with no inherent hydrating activity.

  12. Topical Delivery of Hyaluronic Acid into Skin using SPACE-peptide Carriers

    Science.gov (United States)

    Chen, Ming; Gupta, Vivek; Anselmo, Aaron C.; Muraski, John A.; Mitragotri, Samir

    2014-01-01

    Topical penetration of macromolecules into skin is limited by their low permeability. Here, we report the use of a skin penetrating peptide, SPACE peptide, to enhance topical delivery of a macromolecule, hyaluronic acid (HA, MW: 200–325 kDa). The peptide was conjugated to phospholipids and used to prepare an ethosomal carrier system (~110 nm diameter), encapsulating HA. The SPACE-ethosomal system (SES) enhanced HA penetration into porcine skin in vitro by 7.8+/−1.1-fold compared to PBS. The system also enhanced penetration of HA in human skin in vitro, penetrating deep into the epidermis and dermis in skin of both species. In vivo experiments performed using SKH1 hairless mice also confirmed increased dermal penetration of HA using the delivery system; a 5-fold enhancement in penetration was found compared to PBS control. Concentrations of HA in skin were about 1000-fold higher than those in blood; confirming the localized nature of HA delivery into skin. The SPACE-ethosomal delivery system provides a formulation for topical delivery of macromolecules that are otherwise difficult to deliver into skin. PMID:24129342

  13. Transdermal permeation of drugs with differing lipophilicity: Effect of penetration enhancer camphor.

    Science.gov (United States)

    Xie, Feng; Chai, Jia-Ke; Hu, Quan; Yu, Yong-Hui; Ma, Li; Liu, Ling-Ying; Zhang, Xu-Long; Li, Bai-Ling; Zhang, Dong-Hai

    2016-06-30

    The aim of the present study was to investigate the potential application of (+)-camphor as a penetration enhancer for the transdermal delivery of drugs with differing lipophilicity. The skin irritation of camphor was evaluated by in vitro cytotoxicity assays and in vivo transdermal water loss (TEWL) measurements. A series of model drugs with a wide span of lipophilicity (logP value ranging from 3.80 to -0.95), namely indometacin, lidocaine, aspirin, antipyrine, tegafur and 5-fluorouracil, were tested using in vitro transdermal permeation experiments to assess the penetration-enhancing profile of camphor. Meanwhile, the in vivo skin microdialysis was carried out to further investigate the enhancing effect of camphor on the lipophilic and hydrophilic model drugs (i.e. lidocaine and tegafur). SC (stratum corneum)/vehicle partition coefficient and Fourier transform infrared spectroscopy (FTIR) were performed to probe the regulation action of camphor in the skin permeability barrier. It was found that camphor produced a relatively low skin irritation, compared with the frequently-used and standard penetration enhancer laurocapram. In vitro skin permeation studies showed that camphor could significantly facilitate the transdermal absorption of model drugs with differing lipophilicity, and the penetration-enhancing activities were in a parabola curve going downwards with the drug logP values, which displayed the optimal penetration-enhancing efficiency for the weak lipophilic or hydrophilic drugs (an estimated logP value of 0). In vivo skin microdialysis showed that camphor had a similar penetration behavior on transdermal absorption of model drugs. Meanwhile, the partition of lipophilic drugs into SC was increased after treatment with camphor, and camphor also produced a shift of CH2 vibration of SC lipid to higher wavenumbers and decreased the peak area of the CH2 vibration, probably resulting in the alteration of the skin permeability barrier. This suggests that

  14. Grape skins (Vitis vinifera L.) catalyze the in vitro enzymatic hydroxylation of p-coumaric acid to caffeic acid

    DEFF Research Database (Denmark)

    Arnous, Anis; Meyer, Anne S.

    2009-01-01

    The ability of grape skins to catalyze in vitro conversion of p-coumaric acid to the more potent antioxidant caffeic acid was studied. Addition of different concentrations of p-coumaric to red grape skins (Cabernet Sauvignon) resulted in formation of caffeic acid. This caffeic acid formation (Y......) correlated positively and linearly to p-coumaric acid consumption (X): Y = 0.5 X + 9.5; R 2 = 0.96, P skin concentrations, indicated that the grape skins harboured an o......-hydroxylation activity, proposedly a monophenol- or a flavonoid 3′-monooxygenase activity (EC 1.14.18.1 or EC 1.14.13.21). The K m of this crude o-hydroxylation activity in the red grape skin was 0.5 mM with p-coumaric acid....

  15. Magnetic nanoemulsions as drug delivery system for Foscan ®: Skin permeation and retention in vitro assays for topical application in photodynamic therapy (PDT) of skin cancer

    Science.gov (United States)

    Primo, Fernando L.; Michieleto, Leandro; Rodrigues, Marcilene A. M.; Macaroff, Patrícia P.; Morais, Paulo C.; Lacava, Zulmira G. M.; Bentley, Maria Vitória L. B.; Tedesco, Antonio C.

    2007-04-01

    In this work, we performed the synthesis and in vitro characterization of a new class of drug delivery system (DDS) denominated magnetic nanoemulsion (MNE). The association of colloidal nanoparticles with biocompatible magnetic fluids results in a new DDS for application in photodynamic therapy (PDT) and magnetic hyperthermia treatment. It works in a synergic manner with an expected enhancement in tumor damage after minimum drug doses, based on heat dissipation and/or light photosensitization. For this purpose, we investigated the permeation and retention in vitro model using Foscan ® as a photosensitizer incorporated in MNE using a Franz diffusion cell and a biological skin model in biomimetic conditions.

  16. Magnetic nanoemulsions as drug delivery system for Foscan[reg]: Skin permeation and retention in vitro assays for topical application in photodynamic therapy (PDT) of skin cancer

    Energy Technology Data Exchange (ETDEWEB)

    Primo, Fernando L. [Depto. de Quimica, Universidade de Sao Paulo, FFCLRP-USP, 14040-901, Ribeirao Preto-SP (Brazil); Michieleto, Leandro [Depto. de Quimica, Universidade de Sao Paulo, FFCLRP-USP, 14040-901, Ribeirao Preto-SP (Brazil); Rodrigues, Marcilene A.M. [Depto. de Quimica, Universidade de Sao Paulo, FFCLRP-USP, 14040-901, Ribeirao Preto-SP (Brazil); Macaroff, Patricia P. [Depto. de Quimica, Universidade de Sao Paulo, FFCLRP-USP, 14040-901, Ribeirao Preto-SP (Brazil); Morais, Paulo C. [Instituto de Fisica, Universidade de Brasilia, Nucleo de Fisica Aplicada, 70919-970 Brasilia-DF (Brazil); Lacava, Zulmira G.M. [Instituto de Ciencias Biologicas, Universidade de Brasilia, 70910-900 Brasilia -DF (Brazil); Bentley, Maria Vitoria L.B. [Depto. de Ciencias Farmaceuticas, Universidade de Sao Paulo, FCFRP, 14040-901 Ribeirao Preto-SP (Brazil); Tedesco, Antonio C. [Depto. de Quimica, Universidade de Sao Paulo, FFCLRP-USP, 14040-901, Ribeirao Preto-SP (Brazil)]. E-mail: atedesco@usp.br

    2007-04-15

    In this work, we performed the synthesis and in vitro characterization of a new class of drug delivery system (DDS) denominated magnetic nanoemulsion (MNE). The association of colloidal nanoparticles with biocompatible magnetic fluids results in a new DDS for application in photodynamic therapy (PDT) and magnetic hyperthermia treatment. It works in a synergic manner with an expected enhancement in tumor damage after minimum drug doses, based on heat dissipation and/or light photosensitization. For this purpose, we investigated the permeation and retention in vitro model using Foscan[reg] as a photosensitizer incorporated in MNE using a Franz diffusion cell and a biological skin model in biomimetic conditions.

  17. Growth properties of familial Alzheimer skin fibroblasts during in vitro aging.

    Science.gov (United States)

    Tesco, G; Vergelli, M; Amaducci, L; Sorbi, S

    1993-01-01

    Human diploid fibroblasts undergo replicative senescence in vitro, which is strongly correlated with biological aging in vivo. In order to examine whether features compatible with a systemic premature aging are present in familial Alzheimer's disease (FAD) patients, we investigated the growth characteristics of three skin fibroblast lines from FAD patients and from three sex/age-matched controls at different passages until senescence was reached. A kinetic study of the replicative capacity was performed at different culture times by [3H]-thymidine incorporation and crystal violet staining. Data showed no significant difference between the two groups at any studied passage. The life span of the two types of cultures was also comparable. These results suggest that in familial Alzheimer patients there are not systemic signs of accelerated aging.

  18. In vitro percutaneous permeation and skin accumulation of finasteride using vesicular ethosomal carriers.

    Science.gov (United States)

    Rao, Yuefeng; Zheng, Feiyue; Zhang, Xingguo; Gao, Jianqing; Liang, Wenquan

    2008-01-01

    In order to develop a novel transdermal drug delivery system that facilitates the skin permeation of finasteride encapsulated in novel lipid-based vesicular carriers (ethosomes)finasteride ethosomes were constructed and the morphological characteristics were studied by transmission electron microscopy. The particle size, zeta potential and the entrapment capacity of ethosome were also determined. In contrast to liposomes ethosomes were of more condensed vesicular structure and they were found to be oppositely charged. Ethosomes were found to be more efficient delivery carriers with high encapsulation capacities. In vitro percutaneous permeation experiments demonstrated that the permeation of finasteride through human cadaver skin was significantly increased when ethosomes were used. The finasteride transdermal fluxes from ethosomes containing formulation (1.34 +/- 0.11 microg/cm(2)/h) were 7.4, 3.2 and 2.6 times higher than that of finasteride from aqueous solution, conventional liposomes and hydroethanolic solution respectively (P ethosomes produced a significant (P ethosomes are promising vesicular carriers for enhancing percutaneous absorption of finasteride.

  19. Experimental factors affecting in vitro absorption of six model compounds across porcine skin.

    Science.gov (United States)

    Karadzovska, Daniela; Brooks, James D; Riviere, Jim E

    2012-10-01

    This comparative study evaluated the effect of several experimental variables on the absorption of six model [(14)C]-labeled compounds (caffeine, cortisone, diclofenac sodium, mannitol, salicylic acid, and testosterone) through porcine skin. Using static and flow-through diffusion cells, finite or infinite, saturated or unsaturated doses were applied in one of three vehicles: propylene glycol, water, and ethanol following a full factorial experimental design. The flux of each compound into the receptor phase, with or without bovine serum albumin (BSA), was monitored over 24 h. Levels of radioactivity were also determined in the stratum corneum by tape stripping and in the remaining skin. Apparent permeability coefficients (Kp) and dose absorbed were calculated and compared. The overall results emphasize the importance of experimental design and confirm previous findings that identified dose volume, saturation level and vehicle as the main sources of variation in the in vitro assessment of dermal absorption, whilst diffusion cell model and the presence/absence of BSA in the receptor phase had minimal effect. Although the acquired data do not directly reveal new mechanistic information on dermal absorption, the unique and complete study design has provided a suitable data source for the development of dermal absorption prediction models. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Metrics and clinical relevance of percutaneous penetration and lateral spreading.

    Science.gov (United States)

    Vieille-Petit, Aline; Blickenstaff, Nicholas; Coman, Garrett; Maibach, Howard

    2015-01-01

    Percutaneous penetration of urea in vivo in man has been documented. If urea can penetrate the skin, it may also move laterally. Lateral spreading of topical substances leads to unpredictable penetration dynamics and increased skin surface area exposure. The ability of urea, a low molecular-weight hydrophilic model, to penetrate the stratum corneum (SC) and spread outside the application site was investigated in vitro using tape stripping with spectroscopy. The parameters investigated were the following: time between urea application and tape stripping, formulations containing urea and use of a petrolatum-covered ring barrier around the marked application area. The percentage of urea was determined in and around the application site. The spreading of topically applied urea to neighboring areas occurred and was time but not formulation dependent. A significant difference between protocols with and without the petrolatum ring was observed. These results suggest the clinical importance of lateral spreading, occurring predominately on the skin surface. SC thickness varies between anatomical sites, predisposing areas such as the face and scalp margins to increased percutaneous penetration of topical products. The use of a protective petrolatum ring can inhibit lateral spreading of hair dye in individuals allergic to hair dye, limit systemic absorption and increase accuracy when assessing penetration dynamics. © 2014 S. Karger AG, Basel.

  1. In vivo and in vitro evaluation of topical formulations containing physiological lipid mixture for replacement of skin barrier function.

    Science.gov (United States)

    Barba, C; Parra, J L; Coderch, L; Semenzato, A

    2014-06-01

    The aim of the study was to describe a new in vivo and in vitro approach of the efficacy evaluation of cosmetic emollients to better understand the link between the formulation and the activity of cosmetic products. Two long term in vivo studies were carried out on nine healthy Caucasian volunteers mean age 40±12 years to evaluate the protecting and repairing effects of the two different barrier repair cosmetic formulations. The application of the formulations was repeated once a day during 7 days and biophysical parameters (TEWL and Skin Hydration) were measured before and after Sodium laureth sulphate exposure The in vitro study was carried out by freeze substitution transmission electron microscopy (FSTEM) on stratum corneum samples obtained by sections of fresh skin from young pigs, depleted with a solvent mixture and treated with the two products The in vivo results demonstrated that daily product application provided a reinforcement of the skin barrier with protecting and repairing effects from chemical injuries the extent of which was dependent on the formulation features (product A>product B) The role of the technical form on the lipid availability was confirmed by the in vitro evaluation tests. The results point out that a daily application of physiological lipid mixture containing emulsion can protect healthy skin and promote the reparing effect on unpaired barrier skin, reducing TEWL and maintaining hydration of the stratum corneum. The efficacy degree is higher when the cosmetic form promotes the availability of active ingredients increasing the product performance.

  2. New Enlightenment of Skin Cancer Chemoprevention through Phytochemicals: In Vitro and In Vivo Studies and the Underlying Mechanisms.

    Science.gov (United States)

    Singh, Madhulika; Suman, Shankar; Shukla, Yogeshwer

    2014-01-01

    Skin cancer is still a major cause of morbidity and mortality worldwide. Skin overexposure to ultraviolet irradiations, chemicals, and several viruses has a capability to cause severe skin-related disorders including immunosuppression and skin cancer. These factors act in sequence at various steps of skin carcinogenesis via initiation, promotion, and/or progression. These days cancer chemoprevention is recognized as the most hopeful and novel approach to prevent, inhibit, or reverse the processes of carcinogenesis by intervention with natural products. Phytochemicals have antioxidant, antimutagenic, anticarcinogenic, and carcinogen detoxification capabilities thereby considered as efficient chemopreventive agents. Considerable efforts have been done to identify the phytochemicals which may possibly act on one or several molecular targets that modulate cellular processes such as inflammation, immunity, cell cycle progression, and apoptosis. Till date several phytochemicals in the light of chemoprevention have been studied by using suitable skin carcinogenic in vitro and in vivo models and proven as beneficial for prevention of skin cancer. This revision presents a comprehensive knowledge and the main molecular mechanisms of actions of various phytochemicals in the chemoprevention of skin cancer.

  3. New Enlightenment of Skin Cancer Chemoprevention through Phytochemicals: In Vitro and In Vivo Studies and the Underlying Mechanisms

    Science.gov (United States)

    Singh, Madhulika; Suman, Shankar; Shukla, Yogeshwer

    2014-01-01

    Skin cancer is still a major cause of morbidity and mortality worldwide. Skin overexposure to ultraviolet irradiations, chemicals, and several viruses has a capability to cause severe skin-related disorders including immunosuppression and skin cancer. These factors act in sequence at various steps of skin carcinogenesis via initiation, promotion, and/or progression. These days cancer chemoprevention is recognized as the most hopeful and novel approach to prevent, inhibit, or reverse the processes of carcinogenesis by intervention with natural products. Phytochemicals have antioxidant, antimutagenic, anticarcinogenic, and carcinogen detoxification capabilities thereby considered as efficient chemopreventive agents. Considerable efforts have been done to identify the phytochemicals which may possibly act on one or several molecular targets that modulate cellular processes such as inflammation, immunity, cell cycle progression, and apoptosis. Till date several phytochemicals in the light of chemoprevention have been studied by using suitable skin carcinogenic in vitro and in vivo models and proven as beneficial for prevention of skin cancer. This revision presents a comprehensive knowledge and the main molecular mechanisms of actions of various phytochemicals in the chemoprevention of skin cancer. PMID:24757666

  4. New Enlightenment of Skin Cancer Chemoprevention through Phytochemicals: In Vitro and In Vivo Studies and the Underlying Mechanisms

    Directory of Open Access Journals (Sweden)

    Madhulika Singh

    2014-01-01

    Full Text Available Skin cancer is still a major cause of morbidity and mortality worldwide. Skin overexposure to ultraviolet irradiations, chemicals, and several viruses has a capability to cause severe skin-related disorders including immunosuppression and skin cancer. These factors act in sequence at various steps of skin carcinogenesis via initiation, promotion, and/or progression. These days cancer chemoprevention is recognized as the most hopeful and novel approach to prevent, inhibit, or reverse the processes of carcinogenesis by intervention with natural products. Phytochemicals have antioxidant, antimutagenic, anticarcinogenic, and carcinogen detoxification capabilities thereby considered as efficient chemopreventive agents. Considerable efforts have been done to identify the phytochemicals which may possibly act on one or several molecular targets that modulate cellular processes such as inflammation, immunity, cell cycle progression, and apoptosis. Till date several phytochemicals in the light of chemoprevention have been studied by using suitable skin carcinogenic in vitro and in vivo models and proven as beneficial for prevention of skin cancer. This revision presents a comprehensive knowledge and the main molecular mechanisms of actions of various phytochemicals in the chemoprevention of skin cancer.

  5. Proof of concept testing of a positive reference material for in vivo and in vitro skin irritation testing.

    Science.gov (United States)

    Nomura, Yusuke; Lee, Michelle; Fukui, Chie; Watanabe, Kayo; Olsen, Daniel; Turley, Audrey; Morishita, Yuki; Kawakami, Tsuyoshi; Yuba, Toshiyasu; Fujimaki, Hideo; Inoue, Kaoru; Yoshida, Midori; Ogawa, Kumiko; Haishima, Yuji

    2017-12-11

    In vivo and in vitro irritation testing is important for evaluating the biological safety of medical devices. Here, the performance of positive reference materials for skin irritation testing was evaluated. Four reference standards, referred to as Y-series materials, were analyzed: a polyvinyl chloride (PVC) sheet spiked with 0 (Y-1), 1.0 (Y-2), 1.5 (Y-3), or 10 (Y-4) parts of Genapol X-080 per 100 parts of PVC by weight. Y-1, Y-2, and Y-3 did not induce skin irritation responses in an in vitro reconstructed human epidermis (RhE) tissue model, as measured by tissue viability or interleukin-1α release, or in an in vivo intracutaneous response test using rabbits. In contrast, Y-4 extracts prepared with saline or sesame oil at 37°C and 50°C clearly elicited positive irritation responses, including reduced viability (< 50%) and significantly higher interleukin-1α release compared with the solvent alone group, in the RhE tissue model and an intracutaneous response test, where substantial necrosis was observed by histopathology. The positive skin irritation responses induced in vitro under various extraction conditions, as well as those elicited in vivo, indicate that Y-4 is an effective extractable positive control material for in vivo and in vitro skin irritation tests of medical devices. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

  6. Non-Invasive Assessment of Skin Barrier Properties: Investigating Emerging Tools for In Vitro and In Vivo Applications

    Directory of Open Access Journals (Sweden)

    Emer Duffy

    2017-10-01

    Full Text Available There is increasing interest in the development of non-invasive tools for studying the properties of skin, due to the potential for non-destructive sampling, reduced ethical concerns and the potential comparability of results in vivo and in vitro. The present research focuses on the use of a range of non-invasive approaches for studying skin and skin barrier properties in human skin and human skin equivalents (HSE. Analytical methods used include pH measurements, electrical sensing of the epidermis and detection of volatile metabolic skin products. Standard probe based measurements of pH and the tissue dielectric constant (TDC are used. Two other more novel approaches that utilise wearable platforms are also demonstrated here that can assess the electrical properties of skin and to profile skin volatile species. The potential utility of these wearable tools that permit repeatability of testing and comparability of results is considered through application of our recently reported impedance-based tattoo sensors and volatile samplers on both human participants and HSEs. The HSE exhibited a higher pH (6.5 and TDC (56 than human skin (pH 4.9–5.6, TDC 29–36, and the tattoo sensor revealed a lower impedance signal for HSEs, suggesting the model could maintain homeostasis, but in a different manner to human skin, which demonstrated a more highly resistive barrier. Characterisation of volatiles showed a variety of compound classes emanating from skin, with 16 and 27 compounds identified in HSEs and participants respectively. The continuing development of these tools offers potential for improved quality and relevance of data, and potential for detection of changes that are undetectable in traditional palpable and visual assessments, permitting early detection of irritant reactions.

  7. Ethosomes for skin delivery of ammonium glycyrrhizinate: in vitro percutaneous permeation through human skin and in vivo anti-inflammatory activity on human volunteers.

    Science.gov (United States)

    Paolino, Donatella; Lucania, Giuseppe; Mardente, Domenico; Alhaique, Franco; Fresta, Massimo

    2005-08-18

    The aim of this work was the evaluation of various ethosomal suspensions made up of water, phospholipids and ethanol at various concentrations for their potential application in dermal administration of ammonium glycyrrhizinate, a useful drug for the treatment of various inflammatory-based skin diseases. Physicochemical characterization of ethosomes was carried out by photon correlation spectroscopy and freeze fracture electron microscopy. The percutaneous permeation of ammonium glycyrrhizinate/ethosomes was evaluated in vitro through human stratum corneum and epidermis membranes by using Franz's cells and compared with the permeation profiles of drug solutions either in water or in a water-ethanol mixture. Reflectance spectrophotometry was used as a non-invasive technique to evaluate the carrier toxicity, the drug permeation and the anti-inflammatory activity of ammonium glycyrrhizinate in a model of skin erythema in vivo on human volunteers. Ethosomal suspensions had mean sizes ranging from 350 nm to 100 nm as a function of ethanol and lecithin quantities, i.e., high amounts of ethanol and a low lecithin concentration provided ethosome suspensions with a mean size of approximately 100 nm and a narrow size distribution. In vitro and in vivo experiments were carried out by using an ethosome formulation made up of ethanol 45% (v/v) and lecithin 2% (w/v). The ethosome suspension showed a very good skin tolerability in human volunteers, also when applied for a long period (48 h). Ethosomes elicited an increase of the in vitro percutaneous permeation of both methylnicotinate and ammonium glycyrrhizinate. Ethosomes were able to significantly enhance the anti-inflammatory activity of ammonium glycyrrhizinate compared to the ethanolic or aqueous solutions of this drug. Some in vivo experiments also showed the ability of ethosome to ensure a skin accumulation and a sustained release of the ammonium glycyrrhizinate.

  8. Iron Oxide Nanoparticle-Powered Micro-Optical Coherence Tomography for in Situ Imaging the Penetration and Swelling of Polymeric Microneedles in the Skin.

    Science.gov (United States)

    Seeni, Razina Z; Yu, Xiaojun; Chang, Hao; Chen, Peng; Liu, Linbo; Xu, Chenjie

    2017-06-21

    In recent years, polymeric microneedles (MNs) have attracted keen interests among researchers because of their applicability in transdermal drug delivery and interstitial skin fluid (ISF) extraction. When designing and characterizing such devices, it is critical to monitor their real-time in vitro and in vivo performances to optimize the desired effects, yet most of the existing methods are incapable of such functions. To address this unmet need, we develop a real-time noninvasive imaging methodology by integrating iron oxide (Fe3O4) nanoparticles into polymeric MNs to enhance image contrast for micro-optical coherence tomography (μOCT) imaging. Using the Fe3O4-integrated polystyrene-block-poly(acrylic acid) (PS-b-PAA) MNs as an example, we evaluate the influences of Fe3O4 concentrations on contrast enhancement in μOCT imaging and visualize the real-time swelling process of polymeric MNs in biological samples for the first time. Our results show that a concentration of ∼4-5 wt % Fe3O4 nanoparticles not only helps achieve the best contrast-to-noise ratio in μOCT imaging, which is 10 times higher than that without Fe3O4 nanoparticles in air and hydrogel, but also enables the real-time changes in the profile of MNs to be observed clearly in their swelling process in skin tissues. On the basis of such findings, we utilize the optimized concentration of Fe3O4 nanoparticles to further quantitatively study the swelling kinetics of PS-b-PAA MNs in agarose hydrogel and fresh skin tissues, which lasts ∼20 and ∼30-35 s, respectively. The suitability of such a methodology for enhancing μOCT imaging would greatly facilitate the development and clinical translation of MN-based medical technologies.

  9. In vitro psoriasis models with focus on reconstructed skin models as promising tools in psoriasis research.

    Science.gov (United States)

    Desmet, Eline; Ramadhas, Anesh; Lambert, Jo; Van Gele, Mireille

    2017-06-01

    Psoriasis is a complex chronic immune-mediated inflammatory cutaneous disease associated with the development of inflammatory plaques on the skin. Studies proved that the disease results from a deregulated interplay between skin keratinocytes, immune cells and the environment leading to a persisting inflammatory process modulated by pro-inflammatory cytokines and activation of T cells. However, a major hindrance to study the pathogenesis of psoriasis more in depth and subsequent development of novel therapies is the lack of suitable pre-clinical models mimicking the complex phenotype of this skin disorder. Recent advances in and optimization of three-dimensional skin equivalent models have made them attractive and promising alternatives to the simplistic monolayer cultures, immunological different in vivo models and scarce ex vivo skin explants. Moreover, human skin equivalents are increasing in complexity level to match human biology as closely as possible. Here, we critically review the different types of three-dimensional skin models of psoriasis with relevance to their application potential and advantages over other models. This will guide researchers in choosing the most suitable psoriasis skin model for therapeutic drug testing (including gene therapy via siRNA molecules), or to examine biological features contributing to the pathology of psoriasis. However, the addition of T cells (as recently applied to a de-epidermized dermis-based psoriatic skin model) or other immune cells would make them even more attractive models and broaden their application potential. Eventually, the ultimate goal would be to substitute animal models by three-dimensional psoriatic skin models in the pre-clinical phases of anti-psoriasis candidate drugs. Impact statement The continuous development of novel in vitro models mimicking the psoriasis phenotype is important in the field of psoriasis research, as currently no model exists that completely matches the in vivo psoriasis

  10. Estabilidade e estudo de penetração cutânea in vitro da rutina veiculada em uma emulsão cosmética através de um modelo de biomembrana alternativo Stability and in vitro penetration study of rutin incorporated in a cosmetic emulsion through an alternative model biomembrane

    Directory of Open Access Journals (Sweden)

    André Rolim Baby

    2008-06-01

    employed as antioxidant and to prevent the capillary fragility and, when incorporated in cosmetic emulsions, it must target the action site. In vitro cutaneous penetration studies through human skin is the ideal situation, however, there are difficulties to obtain and to maintain this tissue viability. Among the membrane models, shed snake skin presents itself as pure stratum corneum, providing barrier function similar to human and it is obtained without the animal sacrifice. The objectives of this research were the development and stability evaluation of a cosmetic emulsion containing rutin and propylene glycol (penetration enhancer and the evaluation of rutin in vitro cutaneous penetration and retention from the emulsion, employing an alternative model biomembrane. Emulsion was developed with rutin and propylene glycol, both at 5.0% w/w. Active substance presented on the formulation was quantified by a validated spectrophotometric method at 361.0 nm. Rutin cutaneous penetration and retention was performed in vertical diffusion cells with shed snake skin of Crotalus durissus, as alternative model biomembrane, and distilled water and ethanol 99.5% (1:1, as receptor fluid. The experiment was conducted for six hours, at 37.0 ± 0.5 ºC with constant stirring of 300 rpm. Spectrophotometry at 410.0 nm, previously validated, determined the active substance after cutaneous penetration/retention. Emulsion did not promote rutin cutaneous penetration through C. durissus skin, retaining 0.931 ± 0.0391 mg rutin/mg shed snake skin. The referred formulation was chemically stable for 30 days after stored at 25.0 ± 2.0 ºC, 5.0 ± 0.5 ºC and 45.0 ± 0.5 ºC. In conclusion, it has not been verified the active cutaneous penetration through the model biomembrane, but only its retention on the Crotalus durissus stratum corneum, condition considered stable for 30 days.

  11. Constructing Human Skin Equivalents on Porcine Acellular Peritoneum Extracellular Matrix for In Vitro Irritation Testing.

    Science.gov (United States)

    Tsai, Pei-Chin; Zhang, Zheng; Florek, Charles; Michniak-Kohn, Bozena B

    2016-01-01

    The irritancy of topical products has to be investigated to ensure the safety and compliance. Although several reconstructed human epidermal models have been adopted by the Organization for Economic Cooperation and Development (OECD) to replace in vivo animal irritation testing, these models are based on a single cell type and lack dermal components, which may be insufficient to reflect all of the components of irritation. In our study, we investigated the use of acellular porcine peritoneum extracellular matrix as a substrate to construct full-thickness human skin equivalents (HSEs) for use as irritation screening tool. The acellular peritoneum matrix (APM) exhibited excellent skin cell attachment (>80%) and proliferation for human dermal fibroblasts (HDF) and immortalized human keratinocytes (HaCaT). APM-HSEs based on coculture of HDF and HaCaT were prepared. Increased HDF seeding density up to 5 × 10(4)/cm(2) resulted in APM-HSEs with a thicker and more organized epidermis. The epidermis of APM-HSEs expressed keratin 15, a keratinocyte proliferation marker, and involucrin, a differentiation marker, respectively. To assess the use of APM-HSEs for irritation testing, six proficiency chemicals, including three nonirritants (phosphate-buffered saline, polyethylene glycol 400, and isopropanol) and three irritants (1-bromohexane, heptanol, and sodium dodecyl sulfate) were applied. The APM-HSEs were able to discriminate nonirritants from irritants based on the viability. Levels of cytokines (interleukin [IL]-1α, IL-1ra, IL-6, IL-8, and granulocyte macrophage colony-stimulating factor [GM-CSF]) in these treatment groups further assisted the irritancy ranking. In conclusion, we have developed partially differentiated full-thickness APM-HSEs based on acellular porcine peritoneum matrix, and these APM-HSEs demonstrated utility as an in vitro irritation screening tool.

  12. A broad-spectrum sunscreen prevents UVA radiation-induced gene expression in reconstructed skin in vitro and in human skin in vivo.

    Science.gov (United States)

    Marionnet, Claire; Grether-Beck, Susanne; Seité, Sophie; Marini, Alessandra; Jaenicke, Thomas; Lejeune, François; Bastien, Philippe; Rougier, André; Bernerd, Françoise; Krutmann, Jean

    2011-06-01

    The efficacy of sunscreens to protect against ultraviolet (UV) A radiation is usually assessed by measuring erythema formation and pigmentation. The biological relevance of these endpoints for UVA-induced skin damage, however, is not known. We therefore carried out two complementary studies to determine UVA protection provided by a broad-spectrum sunscreen product at a molecular level by studying UVA radiation-induced gene expression. One study was performed on human reconstructed skin in vitro with a semi-global gene expression analysis of 227 genes in fibroblasts and 244 in keratinocytes. The second one was conducted in vivo in human volunteers and focused on genes involved in oxidative stress response and photo-ageing (haeme oxygenase-1, superoxide dismutase-2, glutathione peroxidase, catalase, matrix metalloproteinase-1). In-vitro UVA radiation induced modulation of genes involved in extracellular matrix homeostasis, oxidative stress, heat shock responses, cell growth, inflammation and epidermal differentiation. Sunscreen pre-application abrogated or significantly reduced these effects, as underlined by unsupervised clustering analysis. The in vivo study confirmed that the sunscreen prevented UVA radiation-induced transcriptional expression of the five studied genes. These findings indicate the high efficacy of a broad-spectrum sunscreen in protecting human skin against UVA-induced gene responses and suggest that this approach is a biologically relevant complement to existing methods. © 2011 John Wiley & Sons A/S.

  13. Development of a new in vitro skin sensitization assay (Epidermal Sensitization Assay; EpiSensA) using reconstructed human epidermis.

    Science.gov (United States)

    Saito, Kazutoshi; Nukada, Yuko; Takenouchi, Osamu; Miyazawa, Masaaki; Sakaguchi, Hitoshi; Nishiyama, Naohiro

    2013-12-01

    Recent changes in regulatory requirements and social views on animal testing have accelerated the development of reliable alternative tests for predicting skin sensitizing potential of chemicals. In this study, we aimed to develop a new in vitro skin sensitization assay using reconstructed human epidermis, RhE model, which is expected to have broader applicability domain rather than existing in vitro assays. Microarray analysis revealed that the expression of five genes (ATF3, DNAJB4, GCLM, HSPA6 and HSPH1) related to cellular stress response were significantly up-regulated in RhE model after 6h treatment with representative skin sensitizers, 1-fluoro-2,4-dinitrobenzene and oxazolone, but not a non-sensitizer, benzalkonium chloride. The predictive performance of five genes was examined with eight skin sensitizers (e.g., cinnamic aldehyde), four non-sensitizers (e.g., sodium lauryl sulfate) and four pre-/pro-haptens (e.g., p-phenylenediamine, isoeugenol). When the positive criteria were set to obtain the highest accuracy with the animal testing (LLNA), ATF3, DNAJB4 and GCLM exhibited a high predictive accuracy (100%, 93.8% and 87.5%, respectively). All tested pre-/pro-haptens were correctly predicted by both ATF3 and DNAJB4. These results suggested that the RhE-based assay, termed epidermal sensitization assay (EpiSensA), could be an useful skin sensitization assay with a broad applicability domain including pre-/pro-haptens. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. The active natural anti-oxidant properties of chamomile, milk thistle, and halophilic bacterial components in human skin in vitro.

    Science.gov (United States)

    Mamalis, Andrew; Nguyen, Duc-Huy; Brody, Neil; Jagdeo, Jared

    2013-07-01

    The number of skin cancers continues to rise, accounting for approximately 40% of all cancers reported in the United States and approximately 9,500 deaths per year. Studies have shown reactive oxygen species (ROS) type free radicals are linked to skin cancer and aging. Therefore, it is important for us to identify agents that have anti-oxidant properties to protect skin against free radical damage. The purpose of this research is to investigate the anti-oxidant properties of bisabolol, silymarin, and ectoin that are components from chamomile, milk thistle, and halophilic bacteria, respectively. We measured the ability of bisabolol, silymarin, and ectoin to modulate the hydrogen peroxide (H2O2)-induced upregulation of ROS free radicals in normal human skin fibroblasts in vitro. Using a flow cytometry-based assay, we demonstrated that varying concentrations of these natural components were able to inhibit upregulation of H2O2-generated free radicals in human skin fibroblasts in vitro. Our results indicate components of chamomile, milk thistle, and halophilic bacteria exhibit anti-oxidant capabilities and warrant further study in clinical trials to characterize their anti-cancer and anti-aging capabilities.

  15. Percutaneous Penetration - Methodological Considerations

    DEFF Research Database (Denmark)

    Holmgaard, Rikke; Benfeldt, Eva; Nielsen, Jesper B

    2014-01-01

    Studies on percutaneous penetration are needed to assess the hazards after unintended occupational skin exposures to industrial products as well as the efficacy after intended consumer exposure to topically applied medicinal or cosmetic products. During recent decades, a number of methods have been...... to the vehicles and solvents used in donor and sampling fluids so that it reflects in-use conditions as closely as possible. Based on available experimental data, mathematical models have been developed to aid predictions of skin penetration. The authors question the general use of the present mathematical models...... in hazard assessment, as they seem to ignore outliers among chemicals as well as the heterogeneity of skin barrier properties and skin conditions within the exposed populations....

  16. In vitro human skin permeation of endoxifen: potential for local transdermal therapy for primary prevention and carcinoma in situ of the breast

    Directory of Open Access Journals (Sweden)

    Lee O

    2011-07-01

    Full Text Available Oukseub Lee1, David Ivancic1, Robert T Chatterton Jr2, Alfred W Rademaker3, Seema A Khan11Department of Surgery, 2Department of Obstetrics/Gynecology, 3Department of Preventive Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL, USAPurpose: Oral tamoxifen, a triphenylethylene (TPE, is useful for breast cancer prevention, but its adverse effects limit acceptance by women. Tamoxifen efficacy is related to its major metabolites 4-hydroxytamoxifen (4-OHT and N-desmethyl-4-hydroxytamoxifen (endoxifen [ENX]. Transdermal delivery of these to the breast may avert the toxicity of oral tamoxifen while maintaining efficacy. We evaluated the relative efficiency of skin permeation of 4-OHT and ENX in vitro, and tested oleic acid (OA as a permeation-enhancer.Methods: 4-OHT, ENX, and estradiol (E2 (0.2 mg/mL of 0.5 µCi 3H/mg were dissolved in 60% ethanol-phosphate buffer, ±OA (0.1%–5%. Permeation through EpiDermTM (Matek Corp, Ashland, MA and split-thickness human skin was calculated based on the amount of the agents recovered from the receiver fluid and skin using liquid scintillation counting over 24 hours.Results: In the EpiDerm model, the absorption of 4-OHT and ENX was 10%–11%; total penetration (TP was 26%–29% at 24 hours and was decreased by OA. In normal human skin, the absorption of 4-OHT and ENX was 0.3%; TP was 2%–4% at 24 hours. The addition of 1% OA improved the permeation of ENX significantly more than that of 4-OHT (P < 0.004; further titration of OA at 0.25%–0.5% further improved the permeation of ENX to a level similar to that of estradiol.Conclusion: The addition of OA to ENX results in a favorable rapid delivery equivalent to that of estradiol, a widely used transdermal hormone. The transdermal delivery of ENX to the breast should be further developed in preclinical and clinical studies.Keywords: endoxifen, breast cancer prevention, human skin, transdermal, oleic acid

  17. Assessing the Impact of Mechanical Damage on Full-Thickness Porcine and Human Skin Using an In Vitro Approach

    Directory of Open Access Journals (Sweden)

    Hinda Dabboue

    2015-01-01

    Full Text Available For most xenobiotics, the rates of percutaneous absorption are limited by diffusion through the horny layer of skin. However, percutaneous absorption of chemicals may seriously increase when the skin is damaged. The aim of this work was to develop an in vitro representative model of mechanically damaged skins. The epidermal barrier was examined following exposure to a razor, a rotating brush, and a microneedle system in comparison to tape-stripping which acted as a reference. Excised full-thickness skins were mounted on a diffusion chamber in order to evaluate the effect of injuries and to mimic physiological conditions. The transepidermal water loss (TEWL was greatly increased when the barrier function was compromised. Measurements were made for all the damaged biopsies and observed histologically by microscopy. On human and porcine skins, the tape-stripping application (0 to 40 times showed a proportional increase in TEWL which highlights the destruction of the stratum corneum. Similar results were obtained for all cosmetic instruments. This is reflected in our study by the nonsignificant difference of the mean TEWL scores between 30 strips and mechanical damage. For a specific appreciation, damaged skins were then selected to qualitatively evaluate the absorption of a chlorogenic acid solution using fluorescence microscopy.

  18. Assessing the Impact of Mechanical Damage on Full-Thickness Porcine and Human Skin Using an In Vitro Approach.

    Science.gov (United States)

    Dabboue, Hinda; Builles, Nicolas; Frouin, Éric; Scott, Dan; Ramos, Jeanne; Marti-Mestres, Gilberte

    2015-01-01

    For most xenobiotics, the rates of percutaneous absorption are limited by diffusion through the horny layer of skin. However, percutaneous absorption of chemicals may seriously increase when the skin is damaged. The aim of this work was to develop an in vitro representative model of mechanically damaged skins. The epidermal barrier was examined following exposure to a razor, a rotating brush, and a microneedle system in comparison to tape-stripping which acted as a reference. Excised full-thickness skins were mounted on a diffusion chamber in order to evaluate the effect of injuries and to mimic physiological conditions. The transepidermal water loss (TEWL) was greatly increased when the barrier function was compromised. Measurements were made for all the damaged biopsies and observed histologically by microscopy. On human and porcine skins, the tape-stripping application (0 to 40 times) showed a proportional increase in TEWL which highlights the destruction of the stratum corneum. Similar results were obtained for all cosmetic instruments. This is reflected in our study by the nonsignificant difference of the mean TEWL scores between 30 strips and mechanical damage. For a specific appreciation, damaged skins were then selected to qualitatively evaluate the absorption of a chlorogenic acid solution using fluorescence microscopy.

  19. Extracellular Matrix Modulates Morphology, Growth, Oxidative Stress Response and Functionality of Human Skin Fibroblasts during Aging In Vitro

    DEFF Research Database (Denmark)

    Jørgensen, Peter; Rattan, Suresh

    2014-01-01

    The Hayflick system of cellular aging and replicative senescence in vitro has been used widely in both basic and applied research in biogerontology. The state of replicative senescence is generally considered to be irreversible, but is modifiable by genetic and environmental manipulations. Some...... recent observations indicate that replicative lifespan, senescence and functionality of cells in vitro can be significantly affected by the quality of the extra cellular matrix (ECM). Following up on those reports, here we show that using the ECM prepared from early passage young cells, partial...... rejuvenation of serially passaged human facial skin fibroblasts was possible in pre-senescent middle-aged cells, but not in fully senescent late passage cells. ECM from young cells improved the appearance, viability, stress tolerance and wound healing ability of skin fibroblasts. Furthermore, young ECM...

  20. In Vitro and In Vivo Measurement of Percutaneous Penetration of Low Molecular Weight Toxins of Military Interest

    Science.gov (United States)

    1989-12-15

    results in the release of toxins. Blooms of P. brevis are responsible for massive fish kills in the Gulf of Mexico and along the Florida coast [1]. The...isolated from several Streptomyces strains (fungus- like bacteria). Teleocidin was isolated from Streptomyces medio - cidicus as a strong skin irritant and

  1. The reconstructed skin model as a new tool for investigating in vitro dermal fillers: increased fibroblast activity by hyaluronic acid.

    Science.gov (United States)

    Girardeau-Hubert, Sarah; Teluob, Séverine; Pageon, Hervé; Asselineau, Daniel

    2015-01-01

    Clinical studies on dermal fillers have essentially focused upon visible improvement of skin quality and any eventual side effects, whereas very little is known about their detailed biological effects. New skin equivalent models were created to investigate the biological impact of hyaluronic acid (HA) fillers on the dermal compartment in vitro. Two different reconstructed skin models were developed to incorporate HA within the collagen fibers. In the mixed model, HA was distributed throughout the whole collagen gel whereas the HA was concentrated in the center of collagen gel in the inclusion model. A comparison of the addition of fillers in two models of reconstructed skin has permitted a better understanding of the biological impact of HA fillers. Protein profiling of supernatants from both models suggested a regulation of MMP-1 secretion by fibroblasts as a function of HA volume, distribution in the dermis and degree of cross-linking. Immunostaining of the inclusion model revealed increased production of type I and III procollagens close to the cross-linked HA. Fibroblasts located in this area showed a fusiform morphology as well as an increase in -smooth actin expression. The observed increase in collagen production may thus result in part from tension in fibroblasts surrounding the cross-linked HA. The inclusion reconstructed skin model, as compared to the mixed model, presented here, appears to be a useful tool for investigating the properties of various fillers in vitro and closer to the in vivo situation; our results show that HA fillers promote in vitro remodeling of the dermis by fibroblasts.

  2. In vitro studies of cutaneous retention of magnetic nanoemulsion loaded with zinc phthalocyanine for synergic use in skin cancer treatment

    Energy Technology Data Exchange (ETDEWEB)

    Primo, Fernando L. [Departamento de Ciencias Farmaceuticas, Faculdade de Ciencias Farmaceuticas de Ribeirao Preto, Universidade de Sao Paulo, Avenida do Cafe, s/n, Ribeirao Preto-SP 14040-903 (Brazil); Rodrigues, Marcilene M.A.; Simioni, Andreza R. [Laboratorio de Fotobiologia e Fotomedicina, Departamento de Quimica, Faculdade de Filosofia, Ciencias e Letras de Ribeirao Preto, Universidade de Sao Paulo, Ribeirao Preto-SP 14040-901 (Brazil); Bentley, Maria V.L.B. [Departamento de Ciencias Farmaceuticas, Faculdade de Ciencias Farmaceuticas de Ribeirao Preto, Universidade de Sao Paulo, Avenida do Cafe, s/n, Ribeirao Preto-SP 14040-903 (Brazil); Morais, Paulo C. [Instituto de Fisica, Nucleo de Fisica Aplicada, Universidade de Brasilia, Brasilia-DF 70919-970 (Brazil); Centro de Nanociencia e Nanobiotecnologia, Universidade de Brasilia, Brasilia-DF 70910-900 (Brazil); Tedesco, Antonio C. [Laboratorio de Fotobiologia e Fotomedicina, Departamento de Quimica, Faculdade de Filosofia, Ciencias e Letras de Ribeirao Preto, Universidade de Sao Paulo, Ribeirao Preto-SP 14040-901 (Brazil); Centro de Nanociencia e Nanobiotecnologia, Universidade de Brasilia, Brasilia-DF 70910-900 (Brazil)], E-mail: atedesco@usp.br

    2008-07-15

    In this study was developed a new nano drug delivery system (NDDS) based on association of biodegradable surfactants with biocompatible magnetic fluid of maguemita citrate derivative. This formulation consists in a magnetic emulsion with nanostructured colloidal particles. Preliminary in vitro experiments showed that the formulation presents a great potential for synergic application in the topical release of photosensitizer drug (PS) and excellent target tissue properties in the photodynamic therapy (PDT) combined with hyperthermia (HPT) protocols. The physical chemistry characterization and in vitro assays were carried out by Zn(II) Phtalocyanine (ZnPc) photosensitizer incorporated into NDDS in the absence and the presence of magnetic fluid, showed good results and high biocompatibility. In vitro experiments were accomplished by tape-stripping protocols for quantification of drug association with different skin tissue layers. This technique is a classical method for analyses of drug release in stratum corneum and epidermis+dermis skin layers. The NDDS formulations were applied directly in pig skin (tissue model) fixed in the cell's Franz device with receptor medium container with a PBS/EtOH 20% solution (10 mM, pH 7.4) at 37 deg. C. After 12 h of topical administration stratum corneum was removed from fifty tapes and the ZnPc retained was evaluated by solvent extraction in dimetil-sulphoxide under ultrasonic bath. These results indicated that magnetic nanoemulsion (MNE) increase the drug release on the deeper skin layers when compared with classical formulation in the absence of magnetic particles. This could be related with the increase of biocompatibility of NDDS due to the great affinity for the polar extracelullar matrix in the skin and also for the increase in the drug partition inside of corneocites wall.

  3. A bioactive peptide analogue for myxoma virus protein with a targeted cytotoxicity for human skin cancer in vitro.

    Science.gov (United States)

    Almansour, Nahlah M; Pirogova, Elena; Coloe, Peter J; Cosic, Irena; Istivan, Taghrid S

    2012-07-17

    Cancer is an international health problem, and the search for effective treatments is still in progress. Peptide therapy is focused on the development of short peptides with strong tumoricidal activity and low toxicity. In this study, we investigated the efficacy of a myxoma virus peptide analogue (RRM-MV) as a candidate for skin cancer therapy. RRM-MV was designed using the Resonant Recognition Model (RRM) and its effect was examined on human skin cancer and normal human skin cells in vitro. Cell cultures were treated with various concentrations of the peptides at different incubation intervals. Cellular morphological changes (apoptosis and necrosis) were evaluated using confocal laser scanning microscopy. The cytotoxic effects of RRM-MV on human skin cancer and normal human skin cells were quantitatively determined by cytotoxicity and cell viability assays. The effect on human erythrocytes was also determined using quantitative hemolysis assay. DNA fragmentation assay was performed to detect early apoptotic events in treated cancer cells. Furthermore, to investigate the possible cell signalling pathway targeted by the peptides treatment, the levels of p-Akt expression in skin cancer and normal cells were detected by immunoblotting. Our results indicate that RRM-MV has a dose-dependent toxic effect on cancer cells only up to 18 h. The immunoblotting results indicated that the RRM-MV slightly increased p-Akt expression in melanoma and carcinoma cells, but did not seem to affect p-Akt expression in normal skin cells. RRM-MV targets and lethally harms cancer cells and leaves normal cells unharmed. It is able to reduce the cancer cell viability, disrupting the LDH activity in cancer cells and can significantly affect cancer progression. Further investigation into other cell signalling pathways is needed in the process leading to the in vivo testing of this peptide to prove its safety as a possible effective treatment for skin cancer.

  4. A bioactive peptide analogue for myxoma virus protein with a targeted cytotoxicity for human skin cancer in vitro

    Directory of Open Access Journals (Sweden)

    Almansour Nahlah M

    2012-07-01

    Full Text Available Abstract Background Cancer is an international health problem, and the search for effective treatments is still in progress. Peptide therapy is focused on the development of short peptides with strong tumoricidal activity and low toxicity. In this study, we investigated the efficacy of a myxoma virus peptide analogue (RRM-MV as a candidate for skin cancer therapy. RRM-MV was designed using the Resonant Recognition Model (RRM and its effect was examined on human skin cancer and normal human skin cells in vitro. Methods Cell cultures were treated with various concentrations of the peptides at different incubation intervals. Cellular morphological changes (apoptosis and necrosis were evaluated using confocal laser scanning microscopy. The cytotoxic effects of RRM-MV on human skin cancer and normal human skin cells were quantitatively determined by cytotoxicity and cell viability assays. The effect on human erythrocytes was also determined using quantitative hemolysis assay. DNA fragmentation assay was performed to detect early apoptotic events in treated cancer cells. Furthermore, to investigate the possible cell signalling pathway targeted by the peptides treatment, the levels of p-Akt expression in skin cancer and normal cells were detected by immunoblotting. Results Our results indicate that RRM-MV has a dose-dependent toxic effect on cancer cells only up to 18 h. The immunoblotting results indicated that the RRM-MV slightly increased p-Akt expression in melanoma and carcinoma cells, but did not seem to affect p-Akt expression in normal skin cells. Conclusions RRM-MV targets and lethally harms cancer cells and leaves normal cells unharmed. It is able to reduce the cancer cell viability, disrupting the LDH activity in cancer cells and can significantly affect cancer progression. Further investigation into other cell signalling pathways is needed in the process leading to the in vivo testing of this peptide to prove its safety as a possible

  5. In vitro evaluation of Spirulina platensis extract incorporated skin cream with its wound healing and antioxidant activities.

    Science.gov (United States)

    Gunes, Seda; Tamburaci, Sedef; Dalay, Meltem Conk; Deliloglu Gurhan, Ismet

    2017-12-01

    Algae have gained importance in cosmeceutical product development due to their beneficial effects on skin health and therapeutical value with bioactive compounds. Spirulina platensis Parachas (Phormidiaceae) is renowned as a potential source of high-value chemicals and recently used in skincare products. This study develops and evaluates skin creams incorporated with bioactive S. platensis extract. Spirulina platensis was cultivated, the aqueous crude extract was prepared and in vitro cytotoxicity of S. platensis extract in the range of 0.001-1% concentrations for 1, 3 and 7 d on HS2 keratinocyte cells was determined. Crude extracts were incorporated in skin cream formulation at 0.01% (w/w) concentration and in vitro wound healing and genotoxicity studies were performed. Immunohistochemical staining was performed to determine the collagen activity. 0.1% S. platensis extract exhibited higher proliferation activity compared with the control group with 198% of cell viability after 3 d. Skin cream including 1.125% S. platensis crude extract showed enhanced wound healing effect on HS2 keratinocyte cell line and the highest HS2 cell viability % was obtained with this concentration. The micronucleus (MN) assay results indicated that S. platensis extract incorporated creams had no genotoxic effect on human peripheral blood cells. Immunohistochemical analysis showed that collagen 1 immunoreactivity was improved by increased extract concentration and it was strongly positive in cells treated with 1.125% extract incorporated skin cream. The cell viability, wound healing activity and genotoxicity results showed that S. platensis incorporated skin cream could be of potential value in cosmeceutical and biomedical applications.

  6. Design and characterisation of a novel in vitro skin diffusion cell system for assessing mass casualty decontamination systems.

    Science.gov (United States)

    Matar, H; Larner, J; Kansagra, S; Atkinson, K L; Skamarauskas, J T; Amlot, R; Chilcott, R P

    2014-06-01

    The efficient removal of contaminants from the outer surfaces of the body can provide an effective means of reducing adverse health effects associated with incidents involving the accidental or deliberate release of hazardous materials. Showering with water is frequently used by first responders as a rapid method of mass casualty decontamination (MCD). However, there is a paucity of data on the generic effectiveness and safety of aqueous decontamination systems. To address these issues, we have developed a new in vitro skin diffusion cell system to model the conditions of a common MCD procedure ("ladder pipe system"). The new diffusion cell design incorporates a showering nozzle, an air sampling port for measurement of vapour loss and/aerosolisation, adjustable (horizontal to vertical) skin orientation and a circulating manifold system (to maintain a specified flow rate, temperature and pressure of shower water). The dermal absorption characteristics of several simulants (Invisible Red S, curcumin and methyl salicylate) measured with the new in vitro model were in good agreement with previous in vitro and in vivo studies. Moreover, these initial studies have indicated that whilst flow rate and water temperature are important factors for MCD, the presence of clothing during showering may (under certain circumstances) cause transfer and spreading of contaminants to the skin surface. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Resistance of acellular dermal matrix materials to microbial penetration.

    Science.gov (United States)

    Fahrenbach, Elizabeth N; Qi, Chao; Ibrahim, Omer; Kim, John Y; Alam, Murad

    2013-05-01

    Acellular dermal matrices have many current and potential applications, but their long-term safety has not been extensively studied. In particular, limited information exists regarding such materials' resistance to infection. To assess the resistance to microbial penetration of common acellular dermal matrix materials used in reconstruction after skin cancer excision, treatment of chronic ulcers and burns, breast reconstruction, hernia repairs, and other applications. Comparative in vitro study of 4 commercially available dermal substitutes for their ability to act as barriers to penetration by common skin pathogens. University-based dermatology and plastic surgery departments and a hospital microbiology laboratory. Four commercially available dermal substitutes, including AlloDerm (LifeCell), FlexHD (Musculoskeletal Transplant Foundation), Strattice (LifeCell), and NeoForm (Mentor Corporation). We tested the 4 dermal matrix materials with the following 4 organisms commonly implicated in wound infections: Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, and Candida albicans. Each material was inoculated with the same concentration of each pathogen. The number of bacterial colonies grown on blood agar plates. AlloDerm and rehydrated FlexHD were found to be the best barriers to penetration by P. aeruginosa. AlloDerm, FlexHD, and Strattice also prevented penetration by S. aureus and S. pyogenes; NeoForm was less effective in withstanding these organisms. The results of this study were inconclusive with regard to C. albicans penetration. Three of the 4 commonly used acellular dermal matrix materials are resistant to in vitro penetration by S. aureus and S. pyogenes and partially resistant to P. aeruginosa. Resistance to fungal pathogens is uncertain. Antimicrobial differences across matrix materials may influence their selection for particular uses, such as treatment of refractory leg ulcers or reconstruction after skin cancer excision.

  8. Influence of beam shape on in-vitro cellular transformations in human skin fibroblasts

    Science.gov (United States)

    Mthunzi, Patience; Forbes, Andrew; Hawkins, Denise; Abrahamse, Heidi; Karsten, Aletta E.

    2005-08-01

    A variety of strategies have been utilised for prevention and treatment of chronic wounds such as leg ulcers, diabetic foot ulcers and pressure sores1. Low Level Laser Therapy (LLLT) has been reported to be an invaluable tool in the enhancement of wound healing through stimulating cell proliferation, accelerating collagen synthesis and increasing ATP synthesis in mitochondria to name but a few2. This study focused on an in-vitro analysis of the cellular responses induced by treatment with three different laser beam profiles namely, the Gaussian (G), Super Gaussian (SG) and Truncated Gaussian (TG), on normal wounded irradiated (WI) and wounded non-irradiated (WNI) human skin fibroblast cells (WS1), to test their influence in wound healing at 632.8 nm using a helium neon (HeNe) laser. For each beam profile, measurements were made using average energy densities over the sample ranging from 0.2 to 1 J, with single exposures on normal wounded cells. The cells were subjected to different post irradiation incubation periods, ranging from 0 to 24 hours to evaluate the duration (time) dependent effects resulting from laser irradiation. The promoted cellular alterations were measured by increase in cell viability, cell proliferation and cytotoxicity. The results obtained showed that treatment with the G compared to the SG and TG beams resulted in a marked increase in cell viability and proliferation. The data also showed that when cells undergo laser irradiation some cellular processes are driven by the peak energy density rather than the energy of the laser beam. We show that there exist threshold values for damage, and suggest optimal operating regimes for laser based wound healing.

  9. In vitro and in vivo percutaneous absorption of retinol from cosmetic formulations: significance of the skin reservoir and prediction of systemic absorption.

    Science.gov (United States)

    Yourick, Jeffrey J; Jung, Connie T; Bronaugh, Robert L

    2008-08-15

    The percutaneous absorption of retinol (Vitamin A) from cosmetic formulations was studied to predict systemic absorption and to understand the significance of the skin reservoir in in vitro absorption studies. Viable skin from fuzzy rat or human subjects was assembled in flow-through diffusion cells for in vitro absorption studies. In vivo absorption studies using fuzzy rats were performed in glass metabolism cages for collection of urine, feces, and body content. Retinol (0.3%) formulations (hydroalcoholic gel and oil-in-water emulsion) containing (3)H-retinol were applied and absorption was measured at 24 or 72 h. All percentages reported are % of applied dose. In vitro studies using human skin and the gel and emulsion vehicles found 0.3 and 1.3% retinol, respectively, in receptor fluid at 24 h. Levels of absorption in the receptor fluid increased over 72 h with the gel and emulsion vehicles. Using the gel vehicle, in vitro rat skin studies found 23% in skin and 6% in receptor fluid at 24 h, while 72-h studies found 18% in skin and 13% in receptor fluid. Thus, significant amounts of retinol remained in rat skin at 24 h and decreased over 72 h, with proportional increases in receptor fluid. In vivo rat studies with the gel found 4% systemic absorption of retinol after 24 h and systemic absorption did not increase at 72 h. Retinol remaining in rat skin after in vivo application was 18% and 13% of the applied dermal dose after 24 and 72 h, respectively. Similar observations were made with the oil-in water emulsion vehicle in the rat. Retinol formed a reservoir in rat skin both in vivo and in vitro. Little additional retinol was bioavailable after 24 h. Comparison of these in vitro and in vivo results for absorption through rat skin indicates that the 24-h in vitro receptor fluid value accurately estimated 24-h in vivo systemic absorption. Therefore, the best single estimate of retinol systemic absorption from in vitro human skin studies is the 24-h receptor fluid

  10. Biodegradable Gelatin Microcarriers Facilitate Re-Epithelialization of Human Cutaneous Wounds - An In Vitro Study in Human Skin.

    Directory of Open Access Journals (Sweden)

    Susanna Lönnqvist

    Full Text Available The possibility to use a suspended tridimensional matrix as scaffolding for re-epithelialization of in vitro cutaneous wounds was investigated with the aid of a human in vitro wound healing model based on viable full thickness skin. Macroporous gelatin microcarriers, CultiSpher-S, were applied to in vitro wounds and cultured for 21 days. Tissue sections showed incorporation of wound edge keratinocytes into the microcarriers and thicker neoepidermis in wounds treated with microcarriers. Thickness of the neoepidermis was measured digitally, using immunohistochemical staining of keratins as epithelial demarcation. Air-lifting of wounds enhanced stratification in control wounds as well as wounds with CultiSpher-S. Immunohistochemical staining revealed expression of keratin 5, keratin 10, and laminin 5 in the neoepidermal component. We conclude that the CultiSpher-S microcarriers can function as tissue guiding scaffold for re-epithelialization of cutaneous wounds.

  11. Biodegradable Gelatin Microcarriers Facilitate Re-Epithelialization of Human Cutaneous Wounds - An In Vitro Study in Human Skin

    Science.gov (United States)

    Lönnqvist, Susanna; Rakar, Jonathan; Briheim, Kristina; Kratz, Gunnar

    2015-01-01

    The possibility to use a suspended tridimensional matrix as scaffolding for re-epithelialization of in vitro cutaneous wounds was investigated with the aid of a human in vitro wound healing model based on viable full thickness skin. Macroporous gelatin microcarriers, CultiSpher-S, were applied to in vitro wounds and cultured for 21 days. Tissue sections showed incorporation of wound edge keratinocytes into the microcarriers and thicker neoepidermis in wounds treated with microcarriers. Thickness of the neoepidermis was measured digitally, using immunohistochemical staining of keratins as epithelial demarcation. Air-lifting of wounds enhanced stratification in control wounds as well as wounds with CultiSpher-S. Immunohistochemical staining revealed expression of keratin 5, keratin 10, and laminin 5 in the neoepidermal component. We conclude that the CultiSpher-S microcarriers can function as tissue guiding scaffold for re-epithelialization of cutaneous wounds. PMID:26061630

  12. Incubation of boar spermatozoa in viscous media by addition of methylcellulose improves sperm quality and penetration rates during in vitro fertilization.

    Science.gov (United States)

    González-Abreu, David; García-Martínez, Soledad; Fernández-Espín, Vanesa; Romar, Raquel; Gadea, Joaquín

    2017-04-01

    This work was designed to study whether viscous media can improve the in vitro sperm functionality in pigs by using methylcellulose as a thickener. Viscosity of porcine oviductal fluid (POF) was compared with culture medium (Tyrode's) supplemented with methylcellulose (MET 0, 0.5 and 1% w/v). Spermatozoa were incubated in the different media (0, 1 and 2 h) and sperm motion parameters, lipid membrane disorder, plasma membrane integrity and reactive oxygen species (ROS) formation were assessed. Fertilization results were assessed i) preincubating spermatozoa in the viscous media followed by gamete coculture in a non-viscous medium; and ii) gamete coculture in the viscous media. Viscosity of POF from early luteal phase was higher than late follicular phase. Medium without methylcellulose presented constant viscosity with increased shear rate, while viscosity of the POF and media with methylcellulose was reduced by increased shear rates. Methylcellulose improved sperm linearity, straightness and the proportion of fast-linear spermatozoa. Moreover, methylcellulose increased the rate of viable spermatozoa with intact acrosome and low lipid disorder, reducing the ROS generation. Preincubation in viscous media increased the penetration rate and the mean number of spermatozoa bound to the zona pellucida (both with 0.5 and 1% MET) and reduced monospermy with 1% MET. On the other hand fertilization in the viscous media reduced penetration rate and increased monospermy. The efficiency of the IVF system was not improved with the use of viscous media. The results show the relevance of increasing viscosity thus making the in vitro media more comparable to physiological conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Fluorescent Penetration Enhancers for Transdermal Applications

    Science.gov (United States)

    Seto, Jennifer E.; Polat, Baris E.; VanVeller, Brett; Lopez, Renata F.V.; Langer, Robert; Blankschtein, Daniel

    2011-01-01

    Chemical penetration enhancers are often used to enhance transdermal drug delivery. However, the fundamental mechanisms that govern the interactions between penetration enhancers and skin are not fully understood. Therefore, the goal of this work was to identify naturally fluorescent penetration enhancers (FPEs) in order to utilize well-established fluorescence techniques to directly study the behavior of FPEs within skin. In this study, 12 fluorescent molecules with amphiphilic characteristics were evaluated as skin penetration enhancers. Eight of the molecules exhibited significant activity as skin penetration enhancers, determined using skin current enhancement ratios. In addition, to illustrate the novel, direct, and non-invasive visualization of the behavior of FPEs within skin, three case studies involving the use of two-photon fluorescence microscopy (TPM) are presented, including visualizing glycerol-mitigated and ultrasound-enhanced FPE skin penetration. Previous TPM studies have indirectly visualized the effect of penetration enhancers on skin by using a fluorescent dye to probe the transdermal pathways of the enhancer. These effects can now be directly visualized and investigated using FPEs. Finally, future studies are proposed for generating FPE design principles. The combination of FPEs with fluorescence techniques represents a useful novel approach for obtaining physical insights on the behavior of penetration enhancers within skin. PMID:22062691

  14. Effects of soap-water wash on human epidermal penetration.

    Science.gov (United States)

    Zhu, Hanjiang; Jung, Eui-Chang; Phuong, Christina; Hui, Xiaoying; Maibach, Howard

    2016-08-01

    Skin decontamination is a primary interventional method used to decrease dermal absorption of hazardous contaminants, including chemical warfare agents, pesticides and industrial pollutants. Soap and water wash, the most common and readily available decontamination system, may enhance percutaneous absorption through the "wash-in effect." To understand better the effect of soap-water wash on percutaneous penetration, and provide insight to improving skin decontamination methods, in vitro human epidermal penetration rates of four C(14) -labeled model chemicals (hydroquinone, clonidine, benzoic acid and paraoxon) were assayed using flow-through diffusion cells. Stratum corneum (SC) absorption rates of these chemicals at various hydration levels (0-295% of the dry SC weights) were determined and compared with the results of the epidermal penetration study to clarify the effect of SC hydration on skin permeability. Results showed accelerated penetration curves of benzoic acid and paraoxon after surface wash at 30 min postdosing. Thirty minutes after washing (60 min postdosing), penetration rates of hydroquinone and benzoic acid decreased due to reduced amounts of chemical on the skin surface and in the SC. At the end of the experiment (90 min postdosing), a soap-water wash resulted in lower hydroquinone penetration, greater paraoxon penetration and similar levels of benzoic acid and clonidine penetration compared to penetration levels in the non-wash groups. The observed wash-in effect agrees with the enhancement effect of SC hydration on the SC chemical absorption rate. These results suggest SC hydration derived from surface wash to be one cause of the wash-in effect. Further, the occurrence of a wash-in effect is dependent on chemical identity and elapsed time between exposure and onset of decontamination. By reducing chemical residue quantity on skin surface and in the SC reservoir, the soap-water wash may decrease the total quantity of chemical absorbed in the

  15. The use of biopartitioning micellar chromatography and immobilized artificial membrane column for in silico and in vitro determination of blood-brain barrier penetration of phenols.

    Science.gov (United States)

    Stępnik, Katarzyna E; Malinowska, Irena

    2013-04-19

    Biopartitioning Micellar Chromatography (BMC) is a mode of micellar liquid chromatography that uses C18 stationary phases and micellar mobile phases of Brij35 under adequate experimental conditions and can be useful to mimic human drug absorption, blood-brain barrier distribution or partitioning processes in biological systems. BMC system can be useful in constructing good predictive models because the characteristics of the BMC system are similar to biological barriers and extracellular fluids. Immobilized Artificial Membrane (IAM) chromatography uses stationary phase which consists of a monolayer of phosphatidylcholine covalently immobilized on an inert silica support. IAM columns are thought to mimic very closely a membrane bilayer and are used in a HPLC system with a physiological buffer as eluent. In this paper the usefulness of BMC and IAM system for in silico and in vitro determination of blood-brain barrier (BBB) penetration of phenols has been demonstrated. The most important pharmacokinetic parameters of brain have been obtained for the determination of BBB penetration, i.e. BBB permeability - surface area product (PS), usually given as a logPS, brain/plasma equilibration rate (log(PS×fu,brain)) and fraction unbound in plasma (Fu). Moreover, the relationships between retention of eighteen phenols and different parameters of molecular size, lipophilicity and BBB penetration were studied. Extrapolated to pure water values of the logarithms of retention factors (logkw) have been compared with the corresponding octanol-water partition coefficient (logPo-w) values of the solutes. In addition, different physicochemical parameters from Foley's equation for BMC system have been collated with the chromatographic data. The Linear Solvation Energy Relationship (LSER) using Abraham model for the describing of phenols penetration across BBB has been used. Four equations were developed as a multiple linear regression using retention data from IAM and BMC system (QRAR

  16. Topical gels of lidocaine HCl using cashew gum and Carbopol 940: preparation and in vitro skin permeation.

    Science.gov (United States)

    Das, Biswarup; Nayak, Amit Kumar; Nanda, Upendranath

    2013-11-01

    The present study was attempted to prepare novel topical gels of 4% lidocaine HCl using cashew gum and Carbopol 940. The prepared gels were evaluated for pH, viscosity, and in vitro skin permeation through excised porcine skin. The pH of these topical gels was found within the range of 5.98-6.06; whereas, the viscosity was found 4.58 × 10(6) to 4.88 × 10(6) cps. The in vitro skin permeation from these gels showed permeation flux range, 851.34 ± 9.16 to 1568.15 ± 14.03 μg/cm(2)/h. The highest permeation flux (1568.15 ± 14.03 μg/cm(2)/h) was observed, when 0.01% menthol was added, which was higher than that of the marketed 4% lidicaine HCl topical gel (1355.41 ± 10.92 μg/cm(2)/h). These topical gels found best-fit with Korsmeyer-Peppas model and almost the super case-II transport mechanism. The stability study revealed that these gels were physically stable without occurrence of syneresis. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Intralaboratory validation of four in vitro assays for the prediction of the skin sensitizing potential of chemicals.

    Science.gov (United States)

    Bauch, Caroline; Kolle, Susanne N; Fabian, Eric; Pachel, Christina; Ramirez, Tzutzuy; Wiench, Benjamin; Wruck, Christoph J; van Ravenzwaay, Bennard; Landsiedel, Robert

    2011-09-01

    Allergic contact dermatitis is induced by repeated skin contact with an allergen. Assessment of the skin sensitizing potential of chemicals, agrochemicals, and especially cosmetic ingredients is currently performed with the use of animals. Animal welfare and EU legislation demand animal-free alternatives reflected in a testing and marketing ban for cosmetic ingredients beginning in 2013. The underlying mechanisms of induction and elicitation of skin sensitization are complex and a chemical needs to comply several properties being skin sensitizing. To account for the multitude of events in the induction of skin sensitization an in vitro test system will consist of a battery of various tests. Currently, we performed intralaboratory validations of four assays addressing three different events during induction of skin sensitization. (1) The Direct Peptide Reactivity Assay (DPRA) according to Gerberick and co-workers (Gerberick et al., 2004) using synthetic peptides and HPLC analysis. (2) Two dendritic cell activation assays based on the dendritic cell like cell lines U-937 and THP-1 and flow cytometric detection of the maturation markers CD54 and/or CD86 (Ashikaga et al., 2006; Python et al., 2007; Sakaguchi et al., 2006). (3) Antioxidant response element (ARE)-dependent gene activity in a HaCaT reporter gene cell line (Emter et al., 2010). We present the results of our intralaboratory validation of these assays with 23 substances of known sensitizing potential. The sensitivity, specificity, and accuracy of the individual tests were obtained by comparison to human epidemiological data as well as to data from animal tests such as the local lymph node assay. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. In vitro efficacy of tavaborole topical solution, 5% after penetration through nail polish on ex vivo human fingernails.

    Science.gov (United States)

    Gupta, Aditya K; Vlahovic, Tracey C; Foley, Kelly A; Lowe, Nicole Gellings; Turner, Rob; Brown, Marc; Hall, Steve

    2018-01-10

    Topical antifungal treatments for onychomycosis are applied to clean, unpolished nails for 48 weeks or longer. Patients often wish to mask their infection with nail polish yet there is no evidence to suggest antifungal efficacy in the presence of nail polish. To determine if tavaborole retains the ability to penetrate the nail plate and inhibit fungal growth in the presence of nail polish. Tavaborole was applied to human fingernails painted with 2 or 4 coats of nail polish, and unpainted nails in an ex vivo model. Nails were mounted on TurChub ® chambers seeded with Trichophyton rubrum and allowed to incubate for 7 days. Antifungal activity was assessed by measuring zones of inhibition. Tavaborole exhibited antifungal activity in all experimental groups. The zones of inhibition of T. rubrum for all experimental groups (2 or 4 coats of polish, unpolished) were greater than infected controls (polished and unpolished), p s  polished nails and kills T. rubrum in this ex vivo model.

  19. Potential of biopartitioning micellar chromatography as an in vitro technique for predicting drug penetration across the blood-brain barrier.

    Science.gov (United States)

    Escuder-Gilabert, L; Molero-Monfort, M; Villanueva-Camañas, R M; Sagrado, S; Medina-Hernández, M J

    2004-08-05

    The blood-brain barrier (BBB) is considered to be the main barrier to drug transport into the central nervous system (CNS). The BBB restricts the passive diffusion of many drugs from blood to brain. The ease with which any particular drug diffuses across the BBB is determined largely by the molecular features of drugs, and it is therefore possible to predict the BBB permeability of a drug from its molecular structure. Biopartitioning micellar chromatography (BMC), a mode of micellar liquid chromatography that uses micellar mobile phases of Brij35 in adequate experimental conditions, can be useful in mimicking the drug partitioning process into biological systems. Retention in BMC depends on the hydrophobicity, electronic and steric properties of drugs. In this paper, the usefulness of BMC for predicting the BBB penetration ability of drugs expressed as the brain/blood distribution coefficient (BB) is demonstrated. A multiple linear regression (MLR) model that relates the BB distribution coefficients data with BMC retention data and total molar charge is proposed. The model is obtained using 44 heterogeneous drugs including, neutral, anionic, and cationic compounds. A comparison with other reported methodologies to predict the BBB permeability is also presented.

  20. Initial Characterization of the Pig Skin Bacteriome and Its Effect on In Vitro Models of Wound Healing.

    Directory of Open Access Journals (Sweden)

    Matthew K McIntyre

    Full Text Available Elucidating the roles and composition of the human skin microbiome has revealed a delicate interplay between resident microbes and wound healing. Evolutionarily speaking, normal cutaneous flora likely has been selected for because it potentiates or, at minimum, does not impede wound healing. While pigs are the gold standard model for wound healing studies, the porcine skin microbiome has not been studied in detail. Herein, we performed 16S rDNA sequencing to characterize the pig skin bacteriome at several anatomical locations. Additionally, we used bacterial conditioned-media with in vitro techniques to examine the paracrine effects of bacterial-derived proteins on human keratinocytes (NHEK and fibroblasts (NHDF. We found that at the phyla level, the pig skin bacteriome is similar to that of humans and largely consists of Firmicutes (55.6%, Bacteroidetes (20.8%, Actinobacteria (13.3%, and Proteobacteria (5.1% however species-level differences between anatomical locations exist. Studies of bacterial supernatant revealed location-dependent effects on NHDF migration and NHEK apoptosis and growth factor release. These results expand the limited knowledge of the cutaneous bacteriome of healthy swine, and suggest that naturally occurring bacterial flora affects wound healing differentially depending on anatomical location. Ultimately, the pig might be considered the best surrogate for not only wound healing studies but also the cutaneous microbiome. This would not only facilitate investigations into the microbiome's role in recovery from injury, but also provide microbial targets for enhancing or accelerating wound healing.

  1. Topical application of probiotics in skin: adhesion, antimicrobial and antibiofilm in vitro assays.

    Science.gov (United States)

    Lopes, E G; Moreira, D A; Gullón, P; Gullón, B; Cardelle-Cobas, A; Tavaria, F K

    2017-02-01

    When skin dysbiosis occurs as a result of skin disorders, probiotics can act as modulators, restoring microbial balance. Several properties of selected probiotics were evaluated so that their topical application could be considered. Adhesion, antimicrobial, quorum sensing and antibiofilm assays were carried out with several probiotic strains and tested against selected skin pathogens. All tested strains displayed significant adhesion to keratin. All lactobacilli with the exception of Lactobacillus delbrueckii, showed antimicrobial activity against skin pathogens, mainly due to organic acid production. Most of them also prevented biofilm formation, but only Propioniferax innocua was able to break down mature biofilms. This study demonstrates that although all tested probiotics adhered to human keratin, they showed limited ability to prevent adhesion of some potential skin pathogens. Most of the tested probiotics successfully prevented biofilm formation, suggesting that they may be successfully used in the future as a complement to conventional therapies in the treatment of a range of skin disorders. The topically used probiotics may be a natural, targeted treatment approach to several skin disorders and a complement to conventional therapies which present many undesirable side effects. © 2016 The Society for Applied Microbiology.

  2. The Effect and Mechanism of Transdermal Penetration Enhancement of Fu's Cupping Therapy: New Physical Penetration Technology for Transdermal Administration with Traditional Chinese Medicine (TCM) Characteristics.

    Science.gov (United States)

    Xie, Wei-Jie; Zhang, Yong-Ping; Xu, Jian; Sun, Xiao-Bo; Yang, Fang-Fang

    2017-03-27

    In this paper, a new type of physical penetration technology for transdermal administration with traditional Chinese medicine (TCM) characteristics is presented. Fu's cupping therapy (FCT), was established and studied using in vitro and in vivo experiments and the penetration effect and mechanism of FCT physical penetration technology was preliminarily discussed. With 1-(4-chlorobenzoyl)-5-methoxy-2-methylindole-3-ylacetic acid (indomethacin, IM) as a model drug, the establishment of high, medium, and low references was completed for the chemical permeation system via in vitro transdermal tests. Furthermore, using chemical penetration enhancers (CPEs) and iontophoresis as references, the percutaneous penetration effect of FCT for IM patches was evaluated using seven species of in vitro diffusion kinetics models and in vitro drug distribution; the IM quantitative analysis method in vivo was established using ultra-performance liquid chromatography-tandem mass spectrometry technology (UPLC-MS/MS), and pharmacokinetic parameters: area under the zero and first moment curves from 0 to last time t (AUC0-t, AUMC0-t), area under the zero and first moment curves from 0 to infinity (AUC0-∞, AUMC0-∞), maximum plasma concentration (Cmax) and mean residence time (MRT), were used as indicators to evaluate the percutaneous penetration effect of FCT in vivo. Additionally, we used the 3(K) factorial design to study the joint synergistic penetration effect on FCT and chemical penetration enhancers. Through scanning electron microscopy (SEM) and transmission electron microscope (TEM) imaging, micro- and ultrastructural changes on the surface of the stratum corneum (SC) were observed to explore the FCT penetration mechanism. In vitro and in vivo skin permeation experiments revealed that both the total cumulative percutaneous amount and in vivo percutaneous absorption amount of IM using FCT were greater than the amount using CPEs and iontophoresis. Firstly, compared with the

  3. Analysis of Reparative Activity of Platelet Lysate: Effect on Cell Monolayer Recovery In Vitro and Skin Wound Healing In Vivo.

    Science.gov (United States)

    Sergeeva, N S; Shanskii, Ya D; Sviridova, I K; Karalkin, P A; Kirsanova, V A; Akhmedova, S A; Kaprin, A D

    2016-11-01

    Platelet lysate prepared from donor platelet concentrate and pooled according to a developed technique stimulates migration of multipotent mesenchymal stromal cells of the human adipose tissue and promotes healing of the monolayer defect in cultures of human fibroblasts and multipotent mesenchymal stromal cells in vitro in concentrations close those of fetal calf serum (5-10%). Lysate of platelets from platelet-rich rat blood plasma stimulated healing of the skin defect by promoting epithelialization and granulation tissue formation. The regenerative properties of platelet lysate in vivo increased with increasing its concentration.

  4. Nanocrystals for dermal penetration enhancement - Effect of concentration and underlying mechanisms using curcumin as model.

    Science.gov (United States)

    Vidlářová, Lucie; Romero, Gregori B; Hanuš, Jaroslav; Štěpánek, František; Müller, Rainer H

    2016-07-01

    Nanocrystals have received considerable attention in dermal application due to their ability to enhance delivery to the skin and overcome bioavailability issues caused by poor water and oil drug solubility. The objective of this study was to investigate the effect of nanocrystals on the mechanism of penetration behavior of curcumin as a model drug. Curcumin nanocrystals were produced by the smartCrystals® process, i.e. bead milling followed by high pressure homogenization. The mean particle size of the curcumin crystals was about 200nm. Stabilization was performed with alkyl polyglycoside surfactants. The distribution of curcumin within the skin was determined in vitro on cross-sections of porcine skin and visualized by fluorescent microscopy. The skin penetration profile was analyzed for the curcumin nanosuspension with decreasing concentrations (2%, 0.2%, 0.02% and 0.002% by weight) and compared to nanocrystals in a viscous hydroxypropylcellulose (HPC) gel. This study demonstrated there was minor difference between low viscous nanosuspension and the gel, but low viscosity seemed to favor skin penetration. Localization of curcumin was observed in the hair follicles, also contributing to skin uptake. Looking at the penetration of curcumin from formulations with decreasing nanocrystal concentration, formulations with 2%, 0.2% and 0.02% showed a similar penetration profile, whereas a significantly weaker fluorescence was observed in the case of a formulation containing 0.002% of curcumin nanocrystals. In this study we have shown that curcumin nanocrystals prepared by the smartCrystal® process are promising carriers in dermal application and furthermore, we identified the ideal concentration of 0.02% nanocrystals in dermal formulations. The comprehensive study of decreasing curcumin concentration in formulations revealed that the saturation solubility (Cs) is not the only determining factor for the penetration. A new mechanism based also on the concentration of the

  5. Antioxidant cosmeto-textiles: skin assessment.

    Science.gov (United States)

    Alonso, Cristina; Martí, Meritxell; Martínez, Vanessa; Rubio, Laia; Parra, José L; Coderch, Luisa

    2013-05-01

    Resveratrol, a natural product, has been reported to have antioxidant activities such as the scavenging of free radicals. This compound could be used in the dermocosmetic field to protect the skin from oxidative stress. In this work, the percutaneous profile of resveratrol in ethanol solutions through pig skin was determinated by an in vitro methodology. The percutaneous absorption of resveratrol was measured and compared with trolox, an analogous of Vitamin E. Both antioxidants were found in all skin sections (stratum corneum, epidermis, and dermis). Besides, the free radical scavenging activity of resveratrol and trolox has been evaluated using DPPH method. The effective dose (ED₅₀) of compounds and DPPH radical inhibition in each skin layer were evaluated. Under the conditions used for these experiments, it can be deduced that resveratrol is more efficient than trolox as an antioxidant, also in the inner skin layers. The cosmeto-textiles with an active substance incorporated into their structure are increasingly used in the cosmetics and pharmaceutical industries. The action of several cosmeto-textiles on the skin was assessed by in vitro and in vivo methodologies. Samples of these cosmeto-textiles were prepared with resveratrol incorporated into cotton and polyamide fabrics. An in vitro percutaneous absorption was used to demonstrate the delivery of the resveratrol from the textile to the different skin layers (stratum corneum, epidermis, and dermis). Additionally, these cosmeto-textiles containing the antioxidant were applied onto the forearms of volunteers to evaluate the textiles' efficacy in skin penetration. The antioxidant's antiradical capacity was evaluated using the DPPH method. Results showed that resveratrol could be detected in the dermis, epidermis, and stratum corneum (SC) by an in vitro percutaneous absorption method and was also detected in the outermost layers of the SC by an in vivo method (stripping). A smaller amount of resveratrol was

  6. Penetration equations

    Energy Technology Data Exchange (ETDEWEB)

    Young, C.W. [Applied Research Associates, Inc., Albuquerque, NM (United States)

    1997-10-01

    In 1967, Sandia National Laboratories published empirical equations to predict penetration into natural earth materials and concrete. Since that time there have been several small changes to the basic equations, and several more additions to the overall technique for predicting penetration into soil, rock, concrete, ice, and frozen soil. The most recent update to the equations was published in 1988, and since that time there have been changes in the equations to better match the expanding data base, especially in concrete penetration. This is a standalone report documenting the latest version of the Young/Sandia penetration equations and related analytical techniques to predict penetration into natural earth materials and concrete. 11 refs., 6 tabs.

  7. Proniosomes for Penetration Enhancement in Transdermal System

    OpenAIRE

    Samita Singla; S. L. HariKumar; Geeta Aggarwal

    2012-01-01

    Over the last few years an inclusive research has been done over provesicular approach for transdermal drug delivery. Skin has a very tough diffusion barrier inhibiting penetration of drug moiety which is rate limiting barrier for penetration of drugs. There are several approaches that deal with penetration enhancement across the skin. Vesicular and provesicular systems are promising amongst them. Vesicular systems including (niosomes, ethosomes, transfersomes and liposomes) are promising sys...

  8. A novel vesicular carrier, transethosome, for enhanced skin delivery of voriconazole: characterization and in vitro/in vivo evaluation.

    Science.gov (United States)

    Song, Chung Kil; Balakrishnan, Prabagar; Shim, Chang-Koo; Chung, Suk-Jae; Chong, Saeho; Kim, Dae-Duk

    2012-04-01

    This study describes a novel carrier, transethosome, for enhanced skin delivery of voriconazole. Transethosomes (TELs) are composed of phospholipid, ethanol, water and edge activator (surfactants) or permeation enhancer (oleic acid). Characterization of the TELs was based on results from recovery, particle size, transmission electron microscopy (TEM), zeta potential and elasticity studies. In addition, skin permeation profile was obtained using static vertical diffusion Franz cells and hairless mouse skin treated with TELs containing 0.3% (w/w) voriconazole, and compared with those of ethosomes (ELs), deformable liposomes (DLs), conventional liposomes (CLs) and control (polyethylene glycol, PG) solutions. The recovery of the studied vesicles was above 90% in all vesicles, as all of them contained ethanol (7-30%). There was no significant difference in the particles size of all vesicles. The TEM study revealed that the TELs were in irregular spherical shape, implying higher fluidity due to perturbed lipid bilayer compared to that of other vesicles which were of spherical shape. The zeta potential of vesicles containing sodium taurocholate or oleic acid showed higher negative value compared to other vesicles. The elasticities of ELs and TELs were much higher than that of CLs and DLs. Moreover, TELs dramatically enhanced the skin permeation of voriconazole compared to the control and other vesicles (p<0.05). Moreover, the TELs enhanced both in vitro and in vivo skin deposition of voriconazole in the dermis/epidermis region compared to DLs, CLs and control. Therefore, based on the current study, the novel carrier TELs could serve as an effective dermal delivery for voriconazole. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Effect of Lipid Composition on In Vitro Release and Skin Deposition of Curcumin Encapsulated Liposomes

    Directory of Open Access Journals (Sweden)

    Geethi Pamunuwa

    2016-01-01

    Full Text Available Liposomal encapsulation improves numerous physiochemical and biological properties of curcumin. The aim of this work was to impart slow release and skin delivery of curcumin via liposomal encapsulation. Liposomes were made using egg yolk phosphatidylcholine as the staple lipid while incorporating polysorbate 80 and stearylamine to prepare hybrid liposomes and positively charged liposomes, respectively. Negatively charged liposomes exhibited the highest encapsulation efficiencies (87.8±4.3% and loading capacities (3.4±0.2%. The sizes of all formulations were about 250 nm, while stearylamine increased the polydispersity index. Positively charged liposomes showed lower degradation temperatures than negatively charged liposomes by 10–15°C, attributable to the presence of stearylamine. The melting temperatures of positively charged liposomes (40–50°C were much higher than those of negatively charged liposomes (14-15°C, which may have affected release and skin deposition behavior of liposomes. The positively charged liposomes exhibited the slowest release of curcumin in phosphate buffered saline (pH 6.8 and the release profiles of all liposomal formulations conformed to the Gompertz model. The negatively charged liposomes facilitated the highest skin deposition of curcumin as revealed by studies conducted using excised pig ear skin. Concisely, positively and negatively charged liposomes were optimal for slow release and skin deposition of curcumin, respectively.

  10. Comparison and validation of an in vitro skin sensitization strategy using a data set of 33 chemical references.

    Science.gov (United States)

    Clouet, Elodie; Kerdine-Römer, Saadia; Ferret, Pierre-Jacques

    2017-12-01

    Allergic contact dermatitis (ACD) is an adverse health effect that develops following repeated exposure to skin sensitizing chemicals. An animal testing ban has been applied in EU, leading to development of reliably predictive non-animal methods. Several in vitro methods have been developed as alternatives but one single non-animal test method is not been sufficient to fully address since the LLNA test ban. Here, we have selected an ITS (Integrated Testing Strategy) for skin sensitization which focuses on three in vitro methods that covered the first three steps of the AOP (DPRA, SENS-IS or h-CLAT). The aim of this study was to compare these three methods due to the WoE approach based on a 2-out-of-3-assessment. The results of 33 references were compared to in vivo data (especially human). We have shown that tested firstly DPRA and SENS-IS have permitted to conclude on 29 of 33 chemicals, whereas DPRA and h-CLAT on 25, and SENS-IS and h-CLAT on 23. With this sequence, DPRA and SENS-IS and then h-CLAT in case of equivocal results, we conclude more quickly by performing fewer tests. Thereby, we have shown that it is better to follow a preferential sequence than testing chemicals simultaneously with these three methods. Copyright © 2016. Published by Elsevier Ltd.

  11. Influence of Cryopreservation Solution on the In Vitro Culture of Skin Tissues Derived from Collared Peccary (Pecari tajacu Linnaeus, 1758).

    Science.gov (United States)

    Borges, Alana A; Lira, Gabriela P O; Nascimento, Lucas E; Queiroz Neta, Luiza B; Santos, Maria V O; Oliveira, Moacir F; Silva, Alexandre R; Pereira, Alexsandra F

    2017-12-07

    Skin vitrification is a promising and alternative tool for the conservation of biodiversity, especially for wild mammals, such as collared peccaries. Several factors can affect the success of this procedure, such as the cryoprotectant solution used. Therefore, this study was carried out to compare the efficiency of various vitrification solutions for recovery of viable cells after in vitro culture of cryopreserved skin tissues derived from the collared peccary, aiming to study the application in biobanking, where cellular use is not immediately required. Then, Dulbecco's modified Eagle's medium (DMEM) composed of 2.2 g/L sodium bicarbonate and 10% fetal bovine serum (FBS) was supplemented with 3.0 M ethylene glycol (EG) or 3.0 M dimethyl sulfoxide (DMSO) or 1.5 M EG plus 1.5 M DMSO with or without sucrose (SUC; 0.25 M) to produce six solutions for solid-surface vitrification. After warming, skin tissues were cultured in vitro and recovered cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity for developing the growth curve and determining the population doubling time (PDT), and viability by Trypan Blue. The vitrification did not alter the ability of the tissues to adhere to the culture dish, as well as the day of all explants with cell growth, subconfluence samples, subconfluence total time, and PDT (p > 0.05). Moreover, independent of the cryoprotectant solution used, the vitrification altered the day of all attached explants (p  0.05). Additionally, for viability after the third passage, only the EG-SUC group maintained the cell quality (88.3%), when compared with the nonvitrified (97.8%, p > 0.05). In conclusion, DMEM with 10% FBS, 3.0 M EG, and 0.25 M sucrose was the most efficient solution for vitrifying collared peccary skin tissues, leading to the in vitro culture of viable cells.

  12. A targeted ultrasound contrast agent carrying gene and cell-penetrating peptide: preparation and gene transfection in vitro.

    Science.gov (United States)

    Ren, Jianli; Zhang, Ping; Tian, Ju; Zhou, Zhiyi; Liu, Xingzhao; Wang, Dong; Wang, Zhigang

    2014-09-01

    Targeted and high efficient gene delivery is a main issue in gene treatment. Taking advantage of ischemic memory target P-selectin and our previous study-synergistic effects of ultrasound-targeted microbubble destruction (UTMD) and TAT peptide on gene transfection, which were characterized by targeted aggregation and high efficient gene transfection, we set up a 'smart' gene delivery system-targeted ultrasound contrast agent (UCA) carrying gene and cell-permeable peptides (CPP). Such UCA had a strong binding force with DNA which was protected from being hydrolysed by nuclease. Moreover, synergistic effects of UTMD and TAT peptide increased gene transfection. Specifically, the UCA were reacted with an ischemic memory target P-selectin overexpressed by ischemic issues (including ischemic heart disease) and loaded with gene and CPP, which enabled targeted localization and gene delivery to ischemic cells overexpressing P-selectin. We demonstrated their targeting affinity for hypoxia human umbilical vein endothelial cell (HUVEC) and gene transfection in vitro. The results of confocal laser scanning microscopy (CLSM) showed that gene and CPP were distributed on the shell of UCA. Red fluorescence was observed on the surface of targeted UCA using immunofluorescent microscopy, which demonstrated that the antibody was successfully connected to the UCA. The targeted UCA was specifically and tightly binded to hypoxia HUVEC, while there were no or little non-targeted UCA binding around hypoxia HUVEC. 24h after transfection, gene transfection efficiency detected by FCM was higher in targeted group than non-targeted group. Overall, the targeted UCA carrying gene and CPP was prepared successfully. It had a strong target binding capacity to hypoxia HUVEC and high efficient gene transfection, which maybe provide a novel strategy for gene therapy. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Penetration depth, concentration and efficiency of transdermal α-arbutin delivery after ultrasound treatment with albumin-shelled microbubbles in mice.

    Science.gov (United States)

    Liao, Ai-Ho; Ma, Wan-Chun; Wang, Chih-Hung; Yeh, Ming-Kung

    2016-09-01

    Recently, the feasibility and effects of using microbubbles (MBs) as an ultrasound (US) contrast agent for enhancing the penetration in transdermal delivery in vivo have been demonstrated, but the mechanism and efficiency are unclear. This study demonstrates the penetration depth, concentration and efficiency of transdermal α-arbutin delivery during 4 weeks after US treatment with MBs in mice. Experimental animals were randomly divided into the following four groups (n = 5 animals per group): (1) penetrating α-arbutin alone (C), (2) US combined with penetrating α-arbutin, (3) US combined with MBs and penetrating α-arbutin, and (4) US combined with diluted MBs and penetrating α-arbutin (UBD). The penetration depths in agarose phantoms and pigskin were 47 and 84% greater for group UBD, respectively, than for group C. The in vitro skin penetration by 2% α-arbutin after 3 h was 83% greater in group UBD than in group C. The degree of in vivo skin whitening (quantified as the luminosity index) in group UBD significantly increased by 25% after 1 week, 34% after 2 weeks, and then stabilized after 3 weeks at 37% in C57BL/6J mice over a 4-week experimental period. Our results indicate that combined treatment with optimal US and MBs can increase skin permeability so as to enhance α-arbutin delivery to inhibit melanogenesis without damaging the skin in mice.

  14. Medical-grade honey kills antibiotic-resistant bacteria in vitro and eradicates skin colonization

    NARCIS (Netherlands)

    Kwakman, Paulus H. S.; van den Akker, Johannes P. C.; Güçlü, Ahmet; Aslami, Hamid; Binnekade, Jan M.; de Boer, Leonie; Boszhard, Laura; Paulus, Frederique; Middelhoek, Pauline; te Velde, Anje A.; Vandenbroucke-Grauls, Christina M. J. E.; Schultz, Marcus J.; Zaat, Sebastian A. J.

    2008-01-01

    BACKGROUND: Antibiotic resistance among microbes urgently necessitates the development of novel antimicrobial agents. Since ancient times, honey has been used successfully for treatment of infected wounds, because of its antibacterial activity. However, large variations in the in vitro antibacterial

  15. Development of In Vitro Isolated Perfused Porcine Skin Flaps for Study of Percutaneous Absorption of Xenobiotics

    Science.gov (United States)

    1987-06-30

    Dunston; the secretarial and administrative assistance of M. Locke ; and the scientific contributions of Drs. V. Scheidt, T. Manning, and L. Dix for...artery in the flap plus that distal skin which is perfused through its dermal-subdermal plexus . Island flaps survive to at least the same length as

  16. In Vitro Construction of Scaffold-Free Bilayered Tissue-Engineered Skin Containing Capillary Networks

    Directory of Open Access Journals (Sweden)

    Yuan Liu

    2013-01-01

    Full Text Available Many types of skin substitutes have been constructed using exogenous materials. Angiogenesis is an important factor for tissue-engineered skin constructs. In this study, we constructed a scaffold-free bilayered tissue-engineered skin containing a capillary network. First, we cocultured dermal fibroblasts with dermal microvascular endothelial cells at a ratio of 2 : 1. A fibrous sheet was formed by the interactions between the fibroblasts and the endothelial cells, and capillary-like structures were observed after 20 days of coculture. Epithelial cells were then seeded on the fibrous sheet to assemble the bilayered tissue. HE staining showed that tissue-engineered skin exhibited a stratified epidermis after 7 days. Immunostaining showed that the epithelium promoted the formation of capillary-like structures. Transmission electron microscopy (TEM analysis showed that the capillary-like structures were typical microblood vessels. ELISA demonstrated that vascularization was promoted by significant upregulation of vascularization associated growth factors due to interactions among the 3 types of cells in the bilayer, as compared to cocultures of fibroblast and endothelial cells and monocultures.

  17. Development and characterisation of an in vitro photomicronucleus test using ex vivo human skin tissue

    NARCIS (Netherlands)

    Reus, A.A.; Meeuwen, R.N.C. van; Vogel, N. de; Maas, W.J.M.; Krul, C.A.M.

    2011-01-01

    Photosafety testing is of concern for the evaluation of personal care products and pharmaceuticals. Current regulatory guidance state that photosafety should be evaluated for compounds that absorb radiation between 290 and 700 nm with relevant exposure in the skin or eyes. However, oversensitivity

  18. Methods to study differences in cell mobility during skin wound healing in vitro

    NARCIS (Netherlands)

    Monsuur, H.N.; Boink, M.A.; Weijers, E.M.; Roffel, S.; Breetveld, M.; Gefen, A.; van den Broek, L.J.; Gibbs, S.

    2016-01-01

    Wound healing events which occur in humans are difficult to study in animals due to differences in skin physiology. Furthermore there are increasing restrictions in Europe for using animals for testing the therapeutic properties of new compounds. Therefore, in line with the 3Rs (reduction,

  19. Stress-induced responses of human skin fibroblasts in vitro reflect human longevity

    NARCIS (Netherlands)

    Dekker, Pim; Maier, Andrea B.; van Heemst, Diana; de Koning-Treurniet, Corine; Blom, Joke; Dirks, Roeland W.; Tanke, Hans J.; Westendorp, Rudi G.J.

    2009-01-01

    Unlike various model organisms, cellular responses to stress have not been related to human longevity. We investigated cellular responses to stress in skin fibroblasts that were isolated from young and very old subjects, and from offspring of nonagenarian siblings and their partners, representatives

  20. Concealing a shiny facial skin appearance by an Aerogel-based formula. In vitro and in vivo studies.

    Science.gov (United States)

    Cassin, Guillaume; Diridollou, Stephane; Flament, Frederic; Adam, Anne-Sophie; Pierre, Patricia; Colomb, Loic; Morancais, Jean-Luc; Qiu, Huixia

    2017-10-10

    To explore, in vitro and in vivo, the potential interest of an Aerogel-based formula, in concealing a naturally shiny facial skin. In vitro, various formulae and ingredients were applied as a thin film onto contrast plates and studied through measuring the shine induced following pump spraying of a mixture of oleic acid and mineral water as a sebum/sweat mix model. In such a test, an Aerogel ingredient led to very positive results. In vivo, two different formulae with various concentrations of Aerogel were randomly tested on half side of the face vs. bare side of Chinese women, under some provocative environmental conditions, known to enhance facial shine. These conditions comprised a normal activity under a hot and highly humid summer time followed -or not- by a hamam session. Both studies included comparative evaluations using a half-face procedure (treated/untreated or vehicle. In the first case, evaluations were quantitatively carried out whereas the second one was based on a quantitative self-evaluations from standardized full-face photographs RESULTS: In vitro, the tested Aerogel, incorporated at 1% or 2% concentration in a common O/W cosmetic emulsion, shows an immediate light scattering effect, thereby masking shine. Such effect appears of much higher amplitude than that of two other tested particulate ingredients (Talc and Perlite). A noticeable remanence of anti-shine effect was confirmed in vivo in extreme conditions. The latter was self-perceived by all participants in the second study. This results is likely related to the super hydrophobic behavior of the Aerogel. As cosmetic ingredient, this new Aerogel appears as a highly promising ingredient for concealing the facial skin shine, a source of complaint from many consumers living in hot and humid regions. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  1. Clinical case seminar: in vivo and in vitro characterization of a novel germline RET mutation associated with low-penetrant nonaggressive familial medullary thyroid carcinoma.

    Science.gov (United States)

    D'Aloiso, Leonardo; Carlomagno, Francesca; Bisceglia, Michele; Anaganti, Suresh; Ferretti, Elisabetta; Verrienti, Antonella; Arturi, Franco; Scarpelli, Daniela; Russo, Diego; Santoro, Massimo; Filetti, Sebastiano

    2006-03-01

    RET mutation analysis provides useful information on the clinical outcome of medullary thyroid carcinomas (MTCs) and the risk of disease in the family members. The objective of this study was to document genotype-phenotype relationships in an Italian family with a novel RET mutation. RET gene alterations were investigated in a patient with unifocal MTC and her relatives. The identified mutation was subjected to in vitro functional testing. Patients included a female proband who developed MTC at age 60, her five children, and three grandchildren. DNA extracted from the blood and the proband's tumor were analyzed for RET alterations. The transforming potential and mitogenic properties of the identified mutation were investigated. A novel heterozygous germline RET mutation at codon 777 (AAC-->AGC, N-->S) (RET/N777S) was identified in the proband and three of her relatives. Two of the latter presented thyroid nodules, but none had MTC or C cell hyperplasia. The proband's MTC was characterized by late onset and limited aggressiveness, with no evidence of regional lymph node or distant metastases 10 yr after total thyroidectomy. This phenotype is consistent with the RET/N777S mutant's low-grade transforming potential and limited activation of RET tyrosine kinase. Our findings indicate that the newly identified RET/N777S mutation is a low-penetrant cause of MTC disease. This phenotype might be less aggressive than that associated with MEN2A of familial MTC, although close clinical follow-up of carriers is essential.

  2. In Vitro Activities of Tedizolid and Linezolid against Gram-Positive Cocci Associated with Acute Bacterial Skin and Skin Structure Infections and Pneumonia.

    Science.gov (United States)

    Chen, Ko-Hung; Huang, Yu-Tsung; Liao, Chun-Hsing; Sheng, Wang-Hui; Hsueh, Po-Ren

    2015-10-01

    Tedizolid is a novel, expanded-spectrum oxazolidinone with potent activity against a wide range of Gram-positive pathogens. A total of 425 isolates of Gram-positive bacteria were obtained consecutively from patients with acute bacterial skin and skin structure infections (ABSSSIs) or pneumonia. These isolates included methicillin-susceptible Staphylococcus aureus (MSSA) (n = 100), methicillin-resistant Staphylococcus aureus (MRSA) (n = 100), Streptococcus pyogenes (n = 50), Streptococcus agalactiae (n = 50), Streptococcus anginosus group (n = 75), Enterococcus faecalis (n = 50), and vancomycin-resistant enterococci (VRE) (Enterococcus faecium) (n = 50). The MICs of tedizolid and linezolid were determined by the agar dilution method. Tedizolid exhibited better in vitro activities than linezolid against MSSA (MIC90s, 0.5 versus 2 μg/ml), MRSA (MIC90s, 0.5 versus 2 μg/ml), S. pyogenes (MIC90s, 0.5 versus 2 μg/ml), S. agalactiae (MIC90s, 0.5 versus 2 μg/ml), Streptococcus anginosus group (MIC90s, 0.5 versus 2 μg/ml), E. faecalis (MIC90s, 0.5 versus 2 μg/ml), and VRE (MIC90s, 0.5 versus 2 μg/ml). The tedizolid MICs against E. faecalis (n = 3) and VRE (n = 2) intermediate to linezolid (MICs, 4 μg/ml) were 1 μg/ml and 0.5 μg/ml, respectively. The tedizolid MIC90s against S. anginosus, S. constellatus, and S. intermedius were 0.5, 1, and 0.5 μg/ml, respectively, and the rates of susceptibility based on the U.S. FDA MIC interpretive breakpoints to the isolates were 16%, 28%, and 72%, respectively. Tedizolid exhibited 2- to 4-fold better in vitro activities than linezolid against a variety of Gram-positive cocci associated with ABSSSIs and pneumonia. The lower susceptibilities of tedizolid against isolates of S. anginosus and S. constellatus than against those of S. intermedius in Taiwan were noted. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. Skin PAMPA: Application in practice

    Directory of Open Access Journals (Sweden)

    Bálint Sinkó

    2015-01-01

    Full Text Available Transdermal drug delivery has been growing extensively in the past decades, therefore new, reliable and cost-effective in vitro models were demanded to support the research and development on this field. Model membrane of PAMPA mimicking skin penetration was first described in 2006, but the need for more bio-mimetic system has been arisen by new industrial tendencies and a bio-relevant system was published in 2012. Since its first publication Skin PAMPA has already been applied by several universities and industrial groups successfully and the first articles, podium and poster presentations have been appeared. The original Skin PAMPA model has been further developed in order to extend its application for formulations. Examples of liquid or semi-solid formulation projects and transdermal patch studies are available beside standard solution applications. The present review demonstrates the different approaches needed for various type of samples, provides examples of applications and practical conclusions for further improvement.

  4. In vitro differentiation of human skin-derived multipotent stromal cells into putative endothelial-like cells

    Directory of Open Access Journals (Sweden)

    Vishnubalaji Radhakrishnan

    2012-01-01

    Full Text Available Abstract Background Multipotent stem cells have been successfully isolated from various tissues and are currently utilized for tissue-engineering and cell-based therapies. Among the many sources, skin has recently emerged as an attractive source for multipotent cells because of its abundance. Recent literature showed that skin stromal cells (SSCs possess mesoderm lineage differentiation potential; however, the endothelial differentiation and angiogenic potential of SSC remains elusive. In our study, SSCs were isolated from human neonatal foreskin (hNFSSCs and adult dermal skin (hADSSCs using explants cultures and were compared with bone marrow (hMSC-TERT and adipose tissue-derived mesenchymal stem cells (hADMSCs for their potential differentiation into osteoblasts, adipocytes, and endothelial cells. Results Concordant with previous studies, both MSCs and SSCs showed similar morphology, surface protein expression, and were able to differentiate into osteoblasts and adipocytes. Using an endothelial induction culture system combined with an in vitro matrigel angiogenesis assay, hNFSSCs and hADSSCs exhibited the highest tube-forming capability, which was similar to those formed by human umbilical vein endothelial cells (HUVEC, with hNFSSCs forming the most tightly packed, longest, and largest diameter tubules among the three cell types. CD146 was highly expressed on hNFSSCs and HUVEC followed by hADSSCs, and hMSC-TERT, while its expression was almost absent on hADMSCs. Similarly, higher vascular density (based on the expression of CD31, CD34, vWF, CD146 and SMA was observed in neonatal skin, followed by adult dermal skin and adipose tissue. Thus, our preliminary data indicated a plausible relationship between vascular densities, and the expression of CD146 on multipotent cells derived from those tissues. Conclusions Our data is the first to demonstrate that human dermal skin stromal cells can be differentiated into endothelial lineage. Hence, SSCs

  5. Solution of the direct problem of induction logging for bed composites with penetration of solution without consideration of the skin effect

    Energy Technology Data Exchange (ETDEWEB)

    Vaksman, K.G.; Afonina, N.M.; Plyusnin, M.I.

    1981-01-01

    Discussed is the solution of the direct problem of induction logging according to the theory of Doll, and the asymptotic formulas are derived for calculating the geometric factors of the beds which permit an increase in the speed and rate of the calculations. A program is described for determining the apparent resistances by any infrared probe in the cross section containing any number of beds with a zone of penetration.

  6. Nanoparticles through the skin: managing conflicting results of inorganic and organic particles in cosmetics and pharmaceutics.

    Science.gov (United States)

    Bolzinger, Marie-Alexandrine; Briançon, Stéphanie; Chevalier, Yves

    2011-01-01

    Toxicity of nanoparticles is a current scientific issue because of the enhanced reactivity of nanomaterials and their possible easy penetration into the body arising from their small size. Because inorganic particles are present in sunscreen cosmetic products, attention has been focused on cutaneous penetration. But organic particles of various sizes are also used in pharmaceutical applications such as skin care and transdermal drug delivery. It appears that organic and inorganic particles penetrate the skin quite differently. The apparent discrepancy is addressed in this review focusing on skin penetration of inorganic sunscreen particles and organic particles for drug delivery. After a short description of the physicochemical properties of these particles, the skin penetration of both types is reviewed with emphasis on the mechanistic issues and the differences that could account for such conflicting results. It appears that investigations by cosmetic and pharmaceutical communities focused on the main issue, i.e., no toxicity in cosmetics and maximum activity of the drug in pharmaceutics. This leaves several fundamental issues as open questions and this does not allow a rigorous comparison between both types of material. While it is claimed that inorganic nanoparticles can only penetrate the outer layer of the skin, it appears that organic submicron particles and even microparticles reach the dermis in an in vitro cell. Besides particle size, the surface chemistry of the particles and the presence of other excipients in the formulations contribute to skin absorption. Copyright © 2011 John Wiley & Sons, Inc.

  7. Isolation, identification, and pathological effects of beach sand bacterial extract on human skin keratinocytes in vitro

    Directory of Open Access Journals (Sweden)

    Fazli Subhan

    2018-01-01

    Full Text Available Background Beaches are recreational spots for people. However, beach sand contains harmful microbes that affect human health, and there are no established methods for either sampling and identifying beach-borne pathogens or managing the quality of beach sand. Method This study was conducted with the aim of improving human safety at beaches and augmenting the quality of the beach experience. Beach sand was used as a resource to isolate bacteria due to its distinctive features and the biodiversity of the beach sand biota. A selected bacterial isolate termed FSRS was identified as Pseudomonas stutzeri using 16S rRNA sequencing and phylogenetic analysis, and the sequence was deposited in the NCBI GenBank database under the accession number MF599548. The isolated P. stutzeri bacterium was cultured in Luria–Bertani growth medium, and a crude extract was prepared using ethyl acetate to examine the potential pathogenic effect of P. stutzeri on human skin. A human skin keratinocyte cell line (HaCaT was used to assess cell adhesion, cell viability, and cell proliferation using a morphological analysis and a WST-1 assay. Result The crude P. stutzeri extract inhibited cell adhesion and decreased cell viability in HaCaT cells. We concluded that the crude extract of P. stutzeri FSRS had a strong pathological effect on human skin cells. Discussion Beach visitors frequently get skin infections, but the exact cause of the infections is yet to be determined. The beach sand bacterium P. stutzeri may, therefore, be responsible for some of the dermatological problems experienced by people visiting the beach.

  8. Assessment of the in vitro dermal irritation potential of cerium, silver, and titanium nanoparticles in a human skin equivalent model.

    Science.gov (United States)

    Miyani, Vivek A; Hughes, Michael F

    2017-06-01

    Metal nanoparticles can potentially contact human skin during their manufacture and use in commercial products. This study examined the potential of metal nanoparticles to elicit irritant contact dermatitis in a human skin equivalent model (HSEM) derived from keratinocytes. Ag (10-100 nm), TiO2 (22-214 nm), and CeO2 (15-40 nm) nanoparticles were studied. The Ag particles were either coated/shelled with silica or capped with citrate or polyvinylpyrrolidone and were in water. The TiO2 and CeO2 particles were suspended in media containing 10% fetal bovine serum. The particles (1 mg/ml) were applied to the epidermal surface of the HSEM. Positive (5% sodium dodecyl sulfate (SDS)) and negative controls (saline or media) were included. After 1-h exposure at 37 °C, the HSEM was washed with saline to remove the nanoparticles. Following a 42-h incubation (37 °C), HSEM viability was assessed using the MTT assay. A test substance is considered a dermal irritant if the HSEM viability is < 50%. The mean viability for the SDS-treated HSEM was 7.8%. The viabilities of the nanoparticle-treated HSEM were 91% or greater. The Ag, TiO2, and CeO2 nanoparticles examined were not dermal irritants under the conditions used in this study. The stratum corneum of the HSEM may limit penetration of metal nanoparticles to induce toxicity.

  9. Antibacterial activity of LauriPure in vitro and on skin of processed broilers.

    Science.gov (United States)

    Studies were conducted to examine the ability of LauriPure to inhibit the growth of Salmonella Typhimurium, Escherichia coli, Staphylococcus simulans, and Listeria innocua isolates recovered from processed broiler carcasses. In vitro studies were conducted using the Bioscreen C Microbiology Reader t...

  10. Oil Body-Bound Oleosin-rhFGF-10: A Novel Drug Delivery System that Improves Skin Penetration to Accelerate Wound Healing and Hair Growth in Mice.

    Science.gov (United States)

    Li, Wenqing; Yang, Jing; Cai, Jingbo; Wang, Hongyu; Tian, Haishan; Huang, Jian; Qiang, Weidong; Zhang, Linbo; Li, Haiyan; Li, Xiaokun; Jiang, Chao

    2017-10-18

    Recombinant human fibroblast growth factor 10 (rhFGF-10) is frequently used to treat patients with skin injuries. It can also promote hair growth. However, the effective application of rhFGF-10 is limited because of its poor stability and transdermal absorption. In this study, polymerase chain reaction (PCR) and Southern blotting were used to identify transgenic safflowers carrying a gene encoding an oleosin-rhFGF-10 fusion protein. The size and structural integrity of oleosin-rhFGF-10 in oil bodies extracted from transgenic safflower seeds was characterized by polyacrylamide gel electrophoresis and western blotting. Oil body extracts containing oleosin-rhFGF-10 were topically applied to mouse skin. The absorption of oleosin-rhFGF-10 was studied by immunohistochemistry. Its efficiency in promoting wound healing and hair regeneration were evaluated in full thickness wounds and hair growth assays. We identified a safflower line that carried the transgene and expressed a 45 kDa oleosin-rhFGF-10 protein. Oil body-bound oleosin-rhFGF-10 was absorbed by the skin with higher efficiency and speed compared with prokaryotically-expressed rhFGF-10. Oleosin-rhFGF-10 also enhanced wound closure and promoted hair growth better than rhFGF-10. The application of oleosin-rhFGF-10 in oil bodies promoted its delivery through the skin, providing a basis for improved therapeutic effects in enhancing wound healing and hair growth.

  11. Carácterísticas histológicas de piel cultivada in vitro Histological Characteristics Of Skin Culture In Vitro

    Directory of Open Access Journals (Sweden)

    Arango

    2009-12-01

    Full Text Available Los injertos de piel cultivados in vitro han sido utilizados tanto en la regeneración de tejidos de áreas cruentas de la piel (úlceras crónicas y quemaduras de diversos grados, como para el tratamiento de genodermatosis. En nuestro medio existe un alto índice de pacientes con úlceras crónicas y un total de 2319 pacientes quemados, en un período de 10 años. El tratamiento convencional de estos pacientes genera estadías de hospitalización prolongadas y costos hospitalarios muy elevados. En este trabajo se establecieron las condiciones para el cultivo y expansión de queratinocitos y fibroblastos humanos, con el propósito de generar un equivalente cutáneo. A su vez, se evaluaron sus características histológicas con el objeto de ofrecer otras opciones de tratamiento. Las células se obtuvieron a partir de piel proveniente de donantes de órganos y de sobrantes de procedimientos quirúrgicos. Se logró un mayor éxito en la obtención de cultivos primarios, con muestras provenientes de donantes menores de 40 años (65%, comparado con los obtenidos de mayores (33%. En el equivalente cutáneo producido con estas células se demostró que los queratinocitos y los fibroblastos, presentan características funcionales, estructurales y morfológicas semejantes a la piel intacta. El equivalente cutáneo además de conservar las características funcionales y estructurales de la piel intacta, presenta otras ventajas en términos de costos, manipulación y estabilidad frente a otros productos similares importados.In vitro skin culture have been used in the regeneration of skin wound (chronic ulcers and burns, and for genodermatosis treatment. In our country there is a high patient number with chronic ulcers and 2319 burned in a period of 10 years. Conventional treatment generates long hospitalization stays and high costs. We established culture conditions of keratinocytes and fibroblasts expansion, to generate a cutaneous substitute in order to

  12. In vitro and in vivo assessment of the effect of Laurus novocanariensis oil and essential oil in human skin.

    Science.gov (United States)

    Viciolle, E; Castilho, P; Rosado, C

    2012-12-01

    Laurus novocanariensis is an endemic plant from the Madeira Island forest that derives a fatty oil, with a strong spicy odour, from its berries that has been used for centuries in traditional medicine to treat skin ailments. This work aimed to investigate the effect of the application of both the oil and its essential oil on normal skin, to assess their safety and potential benefits. Diffusion studies with Franz cells using human epidermal membranes were conducted. The steady-state fluxes of two model molecules through untreated skin were compared with those obtained after a 2-h pre-treatment with either the oil or the essential oil. Additionally, eleven volunteers participated in the in vivo study that was conducted on the forearm and involved daily application of the oil for 5 days. Measurements were performed every day in the treated site with bioengineering methods that measure erythema, irritation and loss of barrier function. Slightly higher steady-state fluxes were observed for both the lipophilic and the hydrophilic molecule when the epidermal membranes were pre-treated. Nevertheless, such differences had no statistical significance, which seems to confirm that neither the oil nor the essential oil impaired the epidermal barrier. Results collected with the Chromameter, the Laser Doppler Flowmeter and the visual scoring are in agreement with those established in the in vitro study. They indicate that the repeated application of the oil did not cause erythema, because the results observed in the first day of the study were maintained throughout the week. Application of the oil did not affect the skin barrier function, because the transepidermal water loss remained constant throughout the study. The stratum corneum hydration was slightly reduced on days 4 and 5. This work shows that both the oil and the essential oil were well tolerated by the skin and did not cause significant barrier impairment or irritation. © 2012 Society of Cosmetic Scientists and the

  13. Percutaneous penetration studies for risk assessment

    DEFF Research Database (Denmark)

    Sartorelli, Vittorio; Andersen, Helle Raun; Angerer, Jürgen

    2000-01-01

    . In order to predict the systemic risk of dermally absorbed chemicals and to enable agencies to set safety standards, data is needed on the rates of percutaneous penetration of important chemicals. Standardization of in vitro tests and comparison of their results with the in vivo data could produce...... internationally accepted penetration rates and/or absorption percentages very useful for regulatory toxicology. The work of the Percutaneous Penetration Subgroup of EC Dermal Exposure Network has been focussed on the standardization and validation of in vitro experiments, necessary to obtain internationally...... accepted penetration rates for regulatory purposes. The members of the Subgroup analyzed the guidelines on percutaneous penetration in vitro studies presented by various organizations and suggested a standardization of in vitro models for percutaneous penetration taking into account their individual...

  14. The percutaneous permeation of a combination of 0.1% octenidine dihydrochloride and 2% 2-phenoxyethanol (octenisept® through skin of different species in vitro

    Directory of Open Access Journals (Sweden)

    Kietzmann Manfred

    2011-08-01

    Full Text Available Abstract Background A water based combination of 0.1% octenidine dihydrochloride and 2% 2 - phenoxyethanol is registered in many European countries as an antiseptic solution (octenisept® for topical treatment with high antimicrobial activity for human use, but octenidine based products have not been registered for veterinary use yet. The aim of the present study was to investigate whether octenidine dihydrochloride or 2 -phenoxyethanol, the two main components of this disinfectant, permeate through animal skin in vitro. Therefore, permeation studies were conducted using Franz-type diffusion cells. 2 ml of the test compound were applied onto 1.77 cm2 split skin of cats, dogs, cows and horses. To simulate wounded skin, cattle skin was treated with adhesive tapes 100 times, as well. Up to an incubation time of 28 hours samples of the acceptor chamber were taken and were analysed by UV-HPLC. Using the method of the external standard, the apparent permeability coefficient, the flux Jmax, and the recovery were calculated. Furthermore, the residues of both components in the skin samples were determined after completion of the diffusion experiment. Results After 28 hours no octenidine dihydrochloride was found in the receptor chamber of intact skin samples, while 2.7% of the topical applied octenidine dihydrochloride permeated through barrier disrupted cattle skin. 2 - phenoxyethanol permeated through all skin samples with the highest permeability in equine, followed by bovine, canine to feline skin. Furthermore, both components were found in the stratum corneum and the dermis of all split skin samples with different amounts in the examined species. Conclusion For 2-phenoxyethanol the systemic impact of the high absorption rate and a potential toxicological risk have to be investigated in further studies. Due to its low absorption rates through the skin, octenidine dihydrochloride is suitable for superficial skin treatment in the examined species.

  15. In vitro release and diffusion studies of promethazine hydrochloride from polymeric dermatological bases using cellulose membrane and hairless mouse skin.

    Science.gov (United States)

    Babar, A; Ray, S D; Patel, N K; Plakogiannis, F M; Gogineni, P

    1999-02-01

    The study was designed to investigate the feasibility of developing a transdermal drug dosage form of promethazine hydrochloride (PMH). The in vitro release and diffusion characteristics of PMH from various dermatological polymeric bases were studied using cellulose membrane and hairless mouse skin as the diffusion barriers. These included polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), cross-linked microcrystalline cellulose, and carboxyl methyl cellulose sodium (Avicel CL-611), and a modified hydrophilic ointment USP. In addition, the effects of several additive ingredients known to enhance the drug release from topical formulations were evaluated. The general rank order for the drug release from these formulations using cellulose membrane was observed to be PEG > HMPC > Avicel CL-611 > hydrophilic ointment base. The inclusion of the additives had little or no effect on the drug diffusion from these bases, except for the hydrophilic ointment formulation containing 15% ethanol, which provided a significant increase in the drug release. However, when these formulations were studied for drug diffusion through the hairless mouse skin, the Avicel CL-611 base containing 15% ethanol exhibited the optimum drug release. The data also revealed that this formulation gave the highest steady-state flux, diffusion, and permeability coefficient values and correlated well with the amount of drug release.

  16. Studying the effectiveness of penetration enhancers to deliver retinol through the stratum cornum by in vivo confocal Raman spectroscopy.

    Science.gov (United States)

    Mélot, Mickaël; Pudney, Paul D A; Williamson, Ann-Marie; Caspers, Peter J; Van Der Pol, Andre; Puppels, Gerwin J

    2009-08-19

    The purpose of this study is to monitor in vivo the effect of chemical penetration enhancers on the delivery of trans-retinol into human skin. Chemical penetration enhancers reversibly alter barrier properties of the SC by disruption of the membrane structures or maximising drug solubility with the skin. So far, most of permeation or penetration experiments are performed in vitro. Raman spectroscopy is uniquely placed to be able to measure biological processes in vivo and this paper shows for the first time that the effect of penetration enhancer on the delivery of trans-retinol can successfully be measured in vivo using this technique. Here, the volar forearm of volunteers was treated with four formulations. One formulation is a highly effective model delivery system identified from ex vivo experiments: trans-retinol in Propylene Glycol (PG)/ethanol, with PG being a well-known and efficient penetration enhancer. The other three formulations are based on 0.3% trans-retinol in Caprylic/Capric Acid Triglyceride (MYRITOL 318), an oil commonly used in skin creams but in two of them a specific penetration enhancer is added. One contains a lipid extractor, Triton X 100, whereas another formulation contains a lipid fluidiser, Oleic Acid. Solutions were applied once and measurements were performed up to 6 h after treatment. Remarkable differences in the delivery of trans-retinol between formulation with or without penetration enhancer can clearly be seen. Moreover, the type of penetration enhancer is also shown to influence the delivery. While using the Oleic Acid, which is a lipid fluidiser, a better delivery of trans-retinol in the skin can be detected. For the first time, the effect of penetration enhancer on the delivery of trans-retinol has been monitored, non invasively in vivo, with time.

  17. In Vitro Chemopreventive Properties of Green Tea, Rooibos and Honeybush Extracts in Skin Cells

    Directory of Open Access Journals (Sweden)

    Tandeka U. Magcwebeba

    2016-11-01

    Full Text Available The chemopreventive properties of the herbal teas rooibos (Aspalathus linearis and honeybush (Cyclopia spp. have been demonstrated on mouse skin in vivo but the underlying mechanisms are not clear. The aim of the current study was to determine the anti-proliferative and pro-apoptotic activity of methanol and aqueous extracts of rooibos and two Cyclopia species in different skin cells, using green tea (Camellia sinensis as a benchmark. Extracts were also characterised for their major individual polyphenols by high performance liquid chromatography and spectroscopically for the total polyphenol (TP groups. The methanol extract of rooibos, containing higher levels of polyphenols than its aqueous extract, displayed similar activity to green tea as it selectively targeted premalignant cells by inhibiting cell proliferation at lower concentrations whilst inducing apoptosis via membrane depolarisation at higher concentrations. Specific roles of the major rooibos dihydrochalcones and flavanol/proanthocyanidin-type (FLAVA compounds are likely to be involved. The aqueous extracts of the Cyclopia species were more active against cell proliferation and at inducing apoptosis which was associated with a higher FLAVA content and a reduced TP/FLAVA ratio. In contrast, their methanol extracts exhibited a cytoprotective effect against apoptosis which was related to their monomeric xanthone and flavanone content. The underlying chemopreventive properties of green tea and the herbal teas appear to be associated with diverse and complex monomeric/polymeric polyphenolic cell interactions.

  18. Drug in adhesive patch of palonosetron: Effect of pressure sensitive adhesive on drug skin permeation and in vitro-in vivo correlation.

    Science.gov (United States)

    Liu, Chao; Hui, Mei; Quan, Peng; Fang, Liang

    2016-09-25

    Palonosetron (PAL) is recommended for the prevention of chemotherapy-induced nausea and vomiting. The aim of this study was to develop a long-acting PAL transdermal patch to improve patient compliance. We were particularly concerned about the effect of pressure sensitive adhesives (PSAs) on PAL skin permeability. Formulation factors including PSAs, backing films and drug loadings were investigated in the in vitro skin permeation study using rabbit skin. Fourier transform infrared spectrometer study and thermal analysis were conducted to investigate the drug-PSA interaction and thermodynamic activity of PSAs, respectively. The results indicated that high drug skin permeation amount was obtained in PSA DURO-TAK(®)87-2516, which had low interaction potential with PAL and high thermodynamic activity. The optimized patch was composed of PAL of 8 %, DURO-TAK(®)87-2516 as PSA, CoTran™ 9700 as backing film and Scotchpak™ 9744 as release liner. The in vitro skin permeation amount of the optimized patch was 734.0±55.8μg/cm(2) during 3-day administration. The absolute bioavailability of the optimized patch was 43 % in rabbit and a good in vitro-in vivo correlation coefficient was obtained (R(2)=0.989). These results indicated the feasibility of PAL transdermal patch in the prevention of chemotherapy-induced nausea and vomiting. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Comparison of skin decontamination efficacy of commercial decontamination products following exposure to VX on human skin.

    Science.gov (United States)

    Thors, L; Koch, M; Wigenstam, E; Koch, B; Hägglund, L; Bucht, A

    2017-08-01

    The decontamination efficacy of four commercially available skin decontamination products following exposure to the nerve agent VX was evaluated in vitro utilizing a diffusion cell and dermatomed human skin. The products included were Reactive Skin Decontamination Lotion (RSDL), the Swedish decontamination powder 104 (PS104), the absorbent Fuller's Earth and the aqueous solution alldecontMED. In addition, various decontamination procedures were assessed to further investigate important mechanisms involved in the specific products, e.g. decontamination removal from skin, physical removal by sponge swabbing and activation of degradation mechanisms. The efficacy of each decontamination product was evaluated 5 or 30 min after dermal application of VX (neat or diluted to 20% in water). The RSDL-lotion was superior in reducing the penetration of VX through human skin, both when exposed as neat agent and when diluted to 20% in water. Swabbing with the RSDL-sponge during 2 min revealed decreased efficacy compared to applying the RSDL-lotion directly on the skin for 30 min. Decontamination with Fuller's Earth and alldecontMED significantly reduced the penetration of neat concentration of VX through human skin. PS104-powder was insufficient for decontamination of VX at both time-points, independently of the skin contact time of PS104. The PS104-slurry (a mixture of PS104-powder and water), slightly improved the decontamination efficacy. Comparing the time-points for initiated decontamination revealed less penetrated VX for RSDL and Fuller's Earth when decontamination was initiated after 5 min compared to 30 min post-exposure, while alldecontMED displayed similar efficacy at both time-points. Decontamination by washing with water only resulted in a significant reduction of penetrated VX when washing was performed 5 min after exposure, but not when decontamination was delayed to 30 min post-exposure of neat VX. In conclusion, early initiated decontamination with the

  20. Preventing false-negatives in the in vitro skin sensitization testing of acid anhydrides using interleukin-8 release assays.

    Science.gov (United States)

    Narita, Kazuto; Vo, Phuc Thi Hong; Yamamoto, Kenta; Kojima, Hajime; Itagaki, Hiroshi

    2017-08-01

    In vitro safety tests may be used as replacements for animal tests owing to their accuracy and high-throughput performance. However, several in vitro skin sensitization tests produce false-negative results such as acid anhydride. Here, we investigated the relationship between false-negative results of acid anhydride and its hydrolysis by aqueous vehicle. Differences in the pattern of hydrolysis for phthalic anhydride (PAH) due to addition of 1 drop of stock solution of PAH in liquid paraffin (LP) dispersion medium and PAH in DMSO were analyzed in a cell-free system. The results showed that use of LP dispersion medium stabilized the concentration of PAH in water over 5min by sustained-release, although almost all PAH converted to phthalic acid in water within 5min using DMSO. Additionally, treatment of THP-1 cells with PAH and phthalic acid using LP dispersion medium for 5min resulted in a 32-fold increase in IL-8 release for PAH as compared with that in the vehicle control. In contrast, for PAH using aqueous vehicle and phthalic acid using LP dispersion medium, there were no significant increases in IL-8 release. Similarly, using LP dispersion medium, trimellitic anhydride significantly increased IL-8 release was observed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. In vitro susceptibility of methicillin-resistant Staphylococcus aureus isolates from skin and soft tissue infections to vancomycin, daptomycin, linezolid and tedizolid

    Directory of Open Access Journals (Sweden)

    Johanna Marcela Vanegas Múnera

    2017-09-01

    Conclusion: In vitro effectiveness of tedizolid was superior for isolates from skin and soft tissue infections in comparison with the other antibiotics evaluated. The above added to its less toxicity, good bioavailability, daily dose and unnecessity of dosage adjustment, make tedizolid in a promising alternative for the treatment of infections caused by MRSA.

  2. Human systemic exposure to a [14C]-para-phenylenediamine- containing oxidative hair dye and correlation with in vitro percutaneous absorption in human or pig skin

    NARCIS (Netherlands)

    Hueber-Becker, F.; Nohynek, G.J.; Meuling, W.J.A.; Benech-Kieffer, F.; Toutain, H.

    2004-01-01

    We investigated the absorption of a commercial [14C]-PPD- containing oxidative dark-shade hair dye in human volunteers as well as in vitro using human or pig ear skin. The hair of eight male volunteers was cut to a standard length, dyed, washed, dried, clipped and collected. Hair, washing water,

  3. Mass transport model through the skin by microencapsulation system.

    Science.gov (United States)

    Carreras, Núria; Alonso, Cristina; Martí, Meritxell; Lis, Manel J

    2015-01-01

    Skin drug delivery can be subdivided into topical and transdermal administration. Transdermal administration can take advantage of chemical and physical strategies that can improve skin permeability and allow drug penetration. In this study, the development of a skin penetration profile was carried out by an in vitro technique for a microencapsulated system of ibuprofen. Release experiments were performed using percutaneous absorption tests to determine the evolution of the principle present in each of the different skin compartments as a function of time. A general kinetic model for a microencapsulated structure as a mass transport system through the skin was applied: [Formula: see text] This model could predict the penetration profile of encapsulated substances through skin from biofunctional textiles as well as estimate the dosage profile of the active principle. The apparent diffusion coefficients found were 1.20 × 10(-7 )cm/s for the stratum corneum and higher for the rest of the skin 6.67 × 10(-6 )cm/s.

  4. Penetration Time Profiles for Two Class 3B Lasers in In Situ Human Achilles at Rest and Stretched.

    Science.gov (United States)

    Bordvik, Daniel Huseby; Haslerud, Sturla; Naterstad, Ingvill Fjell; Lopes-Martins, Rodrigo Alvaro Brandão; Leal Junior, Ernesto Cesar Pinto; Bjordal, Jan Magnus; Joensen, Jon

    2017-10-01

    The majority of studies investigating penetration of laser light are performed in vitro on skin flaps, with measures of immediate penetration depth and energy loss. The aim of this study was to investigate the penetration time profiles for two different lasers used in low-level laser therapy, during 150 sec of exposure both in stretched and relaxed human Achilles in situ. Thirty-four Achilles tendons from 17 healthy volunteers were irradiated by an 810 nm, 200 mW, continuous- and a 904 nm, 60 mW, super-pulsed laser. Irradiation was performed with the Achilles tendons in relaxed and stretched condition. The energy penetrating skin-skin was measured every 30 sec using an optical power meter. The 810 nm laser penetration ability did not differ significantly in relaxed and stretched condition with 0.17% [standard error of the mean (SEM) 0.02] of mean output power (MOP) and 0.02% (SEM 0.004) of MOP, respectively. The 904 nm laser demonstrated a statistical significant (p penetration ability both in relaxed and stretched Achilles from 0.25% (SEM 0.03) to 0.38% (SEM 0.04) of MOP and from 0.05% (SEM 0.01) to 0.13% (SEM 0.01) of MOP, respectively. The penetrated ability differed between lasers and tissue conditions at all measure points (p penetrates relatively more energy than the 810 nm laser in in situ human Achilles. Moreover, penetration from the super-pulsed 904 nm laser increased during exposure time, whereas penetration from the 810 nm laser was constant. In addition, stretching the Achilles causes a higher energy attenuation by the tissue.

  5. In vitro and cellular activities of the selected fruits residues for skin aging treatment

    Directory of Open Access Journals (Sweden)

    NATTAYA LOURITH

    Full Text Available ABSTRACT Peel extracts of litchi and rambutan, and that of tamarind seed coat were investigated in relation to their utility in skin-aging treatments. Standardized extracts of tamarind were significantly (p < 0.05 more efficient at O2 •- scavenging (IC50 = 27.44 ± 0.09 than those of litchi and rambutan (IC50 = 29.57 ± 0.30 and 39.49 ± 0.52 μg/ml, respectively and the quercetin standard (IC50 = 31.88 ± 0.15 μg/ml. Litchi extract proved significantly (p < 0.05 more effective for elastase and collagenase inhibition (88.29 ± 0.25% and 79.46 ± 0.92%, respectively than tamarind (35.43 ± 0.68% and 57.69 ± 5.97% or rambutan (31.08 ± 0.38% and 53.99 ± 6.18%. All extracts were safe to human skin fibroblasts and inhibit MMP-2, with litchi extract showing significantly (p < 0.01 enhanced inhibition over the standard, vitamin C (23.75 ± 2.74% and 10.42 ± 5.91% at 0.05 mg/ml, respectively. Extracts suppress melanin production in B16F10 melanoma cells through inhibition of tyrosinase and TRP-2, with litchi extract being the most potent, even more so than kojic acid (standard. These results highlight the potential for adding value to agro-industrial waste, as the basis for the sustainable production of innovative, safe, anti-aging cosmetic products.

  6. Efeito do pH na calcificação in vitro de pele porcina In Vitro calcification of porcine skin: influence of pH

    Directory of Open Access Journals (Sweden)

    Thelma M. Batista

    2007-12-01

    Full Text Available A engenharia de tecidos tem sido utilizada como alternativa na reconstrução de tecidos duros e moles. Este estudo teve como objetivo a calcificação "in vitro" de pele porcina visando à obtenção de um material para regeneração de tecido duro. As matrizes de pele porcina foram calcificadas em cela dupla termostatizada a 37 °C em pH 7,4 e pH 9,0 e caracterizadas por microscopia eletrônica de varredura (MEV, termogravimetria (TGA, espectroscopia no infravermelho (FTIR, calorimetria exploratória diferencial (DSC e difração de raios X. Os resultados obtidos por DSC mostraram que as amostras calcificadas têm um pequeno aumento nos valores de temperatura de desnaturação em relação à amostra não calcificada, enquanto as curvas termogravimétricas mostraram uma porcentagem maior de material inorgânico para o pH 7,4 em comparação com as amostras obtidas em pH 9,0. A formação de sais de fosfato de cálcio nas fibras de colágeno foi confirmada por difração de raios X (DRX, espectroscopia no infravermelho (FTIR e microscopia eletrônica de varredura (MEV.Tissue engineering has been used as an alternative in the reconstruction of hard and soft tissues. The objective of this work was to study the in vitro calcification of porcine skin aiming at obtaining a material to be used for regeneration of hard tissue. Pig skin was calcified at 37 °C, pH 7.4 and pH 9.0 and characterized by scanning electron microscopy (SEM, thermogravimetric analysis (TGA, infrared spectroscopy (FTIR, differential scanning calorimetry (DSC and X ray diffraction (XRD. The results obtained by DSC showed that calcified samples had a small increase in the values of denaturation temperature when compared with uncalcified samples. Thermogravimetric curves showed higher quantity of inorganic material for the pH 7.4 matrix when compared to samples obtained in pH 9.0. The formation of calcium phosphate salts on collagen fibers was seen by SEM, which was also confirmed

  7. Efinaconazole 10% and Tavaborole 5% Penetrate Across Poly-ureaurethane 16%: Results of In Vitro Release Testing and Clinical Implications of Onychodystrophy in Onychomycosis.

    Science.gov (United States)

    Adigun, Chris G; Vlahovic, Tracey C; McClellan, Michael B; Thakker, Kailas D; Klein, Ryan R; Elstrom, Tuan A; Ward, Daniel B

    2016-09-01

    Poly-ureaurethane has been previously described for the management of dry, brittle, and in general, dystrophic nails. The polymer yields a waterproof, breathable barrier to protect the nail plate and prevent further damage to the nail, while regulating transonychial water loss (TOWL). Because nail dystrophy and dessication are contributing factors to onychomycosis, a barrier that protects the nail but also allows a topical antifungal to permeate its shield is potentially an advantageous combination. Oral antifungals such as terbinafine, itraconazole, and fluconazole, as well as the newer topical antifungals efinaconazole and tavaborole (although formulated to penetrate the nail unit and work with the porosity and inherent electrical charge of the nail plate), do not take into account nail damage that has been created from years of harboring a dermatophyte infection. Up to 50% of cases presumed to be onychomycosis are in fact onychodystrophy without fungal infection, and laboratory testing for fungus should be obtained prior to initiating antifungal treatment. Whether a nail has onychomycosis, or onychodystrophy due to other causes, barrier function and structural integrity are compromised in diseased nails, and should be addressed. A poly-ureaurethane barrier that protects against wetting/drying, fungal reservoirs, and microtrauma, followed by the addition of oral or topical antifungals after laboratory fungal confirmation may optimize outcomes in the treatment of onychomycosis. The purpose of this work was to determine through in vitro release testing (IVRT) whether poly-ureaurethane 16% allows for penetration of efinaconazole 10% or tavaborole 5%. Results could spur subsequent clinical studies which would have implications for the addition of an antifungal based on fungal confirmation, after addresssing the underlying nail dystrophy primarily. A vertical diffusion cell system was used to evaluate the ability of efinaconazole 10% and tavaborole 5% to penetrate

  8. Penetrating Fire Extinguisher

    Science.gov (United States)

    1985-01-01

    When Feecon Corporation, a manufacturer of fire protection systems, needed a piercing nozzle for larger aircraft, they were assisted by Kennedy Space Center who provided the company with a fire extinguisher with a hard pointed tip that had been developed in case of an orbiter crash landing. The nozzle can penetrate metal skins of aircraft, trains, etc. Feecon obtained a license and now markets its cobra ram piercing nozzle to airport firefighters. Its primary advantage is that the nozzle can be held in one spot during repeated blows of the ram. *This product has been discontinued and is no longer commercially available.

  9. Effect of skin metabolism on dermal delivery of testosterone: qualitative assessment using a new short-term skin model.

    Science.gov (United States)

    Jacques, C; Perdu, E; Jamin, E L; Cravedi, J P; Mavon, A; Duplan, H; Zalko, D

    2014-01-01

    The skin is a metabolically active organ expressing biotransformation enzymes able to metabolize both endogenous molecules and xenobiotics. We investigated the impact of metabolism on the delivery of testosterone through the skin with an ex vivo pig ear skin system as an alternative model for human skin. Penetration, absorption and metabolic capabilities were investigated up to 72 h after application of [(14)C]-testosterone doses of 50-800 nmol on either fresh or frozen skin, with the latter model being metabolically inactive. Testosterone absorption and metabolite production were monitored by radio-HPLC and gas chromatography-mass spectrometry. Testosterone absorption through frozen skin was much lower, irrespective of the dose of testosterone applied, compared to fresh skin. Using fresh skin samples, >95% of the radioactivity recovered in culture media, as well as the skin itself, corresponded to metabolites. These results were compared with the metabolic data obtained from other in vitro systems (liver and skin microsomes). The present work leads to the conclusion that most of the enzymatic activities expressed in liver fractions are also expressed in pig and human skin. The metabolic activity of the skin can modulate the biological activity of pharmaceuticals (and xenobiotics). Consequently, it can also greatly affect transdermal drug delivery.

  10. Chemical Penetration Enhancers for Transdermal Drug Delivery ...

    African Journals Online (AJOL)

    for transdermal administration. The permeation of drug through skin can be enhanced by both chemical penetration enhancement and physical methods. In this review, we have discussed the chemical penetration enhancement technology for transdermal drug delivery as well as the probable mechanisms of action.

  11. Comparison of the cutaneous iontophoretic delivery of rasagiline and selegiline across porcine and human skin in vitro.

    Science.gov (United States)

    Kalaria, Dhaval R; Patel, Pratik; Patravale, Vandana; Kalia, Yogeshvar N

    2012-11-15

    The objective was to investigate the anodal iontophoresis of the MAO-B inhibitors rasagiline (RAS) and selegiline (SEL) across porcine and human skin in vitro. Passive delivery of RAS and SEL from aqueous solution was minimal; however, increasing current density from 0.1 to 0.3 and 0.5 mA/cm(2) produced a linear increase in steady-state iontophoretic flux (J(ss,RAS)=49.1i(d)+27.9 (r(2)=0.96) and J(ss,SEL)=27.8i(d)+25.8 (r(2)=0.98)). In the absence of background electrolyte, a four-fold change in donor concentration (10, 20 and 40 mM) did not produce a statistically significant increase in cumulative permeation of either drug after iontophoresis at 0.5mA/cm(2) for 7h. Co-iontophoresis of acetaminophen confirmed that electromigration was the dominant transport mechanism for both drugs (∼90%). Total iontophoretic delivery of RAS and SEL across porcine and human skin in vitro was statistically equivalent (RAS: 1512.7 ± 163.7 and 1523.6 ± 195.9 μg/cm(2), respectively, and SEL: 1268.7 ± 231.2 and 1298.3 ± 253.3 μg/cm(2), respectively). Transport efficiencies for RAS and SEL were good (ranged from 6.81 to 8.50 and 2.86 to 3.61%, respectively). Furthermore, the delivery efficiency, i.e., the fraction of the drug in the formulation that was delivered was very high (>56% at 0.5 mA/cm(2)). Cumulative permeation of RAS and SEL from carbopol gels, potential drug reservoirs for iontophoretic systems, was 891.5 ± 148.3 and 626.6 ± 162.4 μg/cm(2), respectively; this was less than from solution and was tentatively attributed to either different partitioning or slower drug diffusion in the gel matrix. The results demonstrated that therapeutic amounts of rasagiline and selegiline could be easily delivered by transdermal iontophoresis with simple gel patches of modest surface area. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. In vitro construction of tissue engineered skin for wound repair after escharectomy of third degree scald: An experimental study

    Directory of Open Access Journals (Sweden)

    Zhong-feng MA

    2016-01-01

    Full Text Available Objective  To observe the practicability and effect of tissue engineered skin for repairing the wound after escharectomy of third degree scald (TDSE in rat model. Methods  Epithelial cells and fibroblasts from newborn SD rats were isolated by enzyme digestion method and cultured in vitro, and porcine acellular dermal matrix (PADM without cytotoxicity was prepared by hyperosmotic saline/sodium hydroxide method. The fibroblasts were mixed with bovine type Ⅰ collagen and inoculated on the surface of PADM. Third passage of cultured epidermal cells from newborn SD rats were inoculated on the collagen surface of the dermal matrix to obtain tissue engineered skin, and it was used to prepare epidermal cell sheet. Forty-eight SD rats with TDSE wound were randomly divided into two groups, then tissue engineered skin (experiment group, and epidermal cell sheet (control group graftings were performed to cover the wounds respectively. Finally, gross observation and histological changes were observed in grafted area. The wound healing rate and wound contraction rate were compared between the two groups. Microvessel count (MVC was performed with antiCD34 monoclonal antibody immunohistochemical staining technique, and vascular endothelial cells were labeled. Basal membrane of the skin was identified by immunohistochemical anti-Laminin staining technique. Results  There was no obvious sign of acute rejection of the graft in both groups. The graft survival rate was 75.05%±3.69%, 83.12%±3.13% and 92.03%±3.87% at the 2th, 4th and 6th week respectively in the experimental group. The graft survival rate was 77.63%±3.23%, 83.17%±3.92% and 91.09%±3.35% at the 2th, 4th and 6th week in the control group. There was no significant difference between the two groups (P>0.05, but the contraction rate of the grafts was 9.13%±2.27%, 18.52%±3.40%, 23.92%±3.01% at the 2th, 4th, 6th week, respectively, in the experimental group, and 14.21%±3.05%, 29.12%±3

  13. Lamellar Liquid Crystal Improves the Skin Retention of 3-O-Ethyl-Ascorbic Acid and Potassium 4-Methoxysalicylate In Vitro and In Vivo for Topical Preparation.

    Science.gov (United States)

    Li, Yuanru; Dong, Cuilian; Cun, Dongmei; Liu, Jie; Xiang, Rongwu; Fang, Liang

    2016-06-01

    The study aimed at increasing the skin retention of 3-O-ethyl-ascorbic acid (EA) and potassium 4-methoxysalicylate (4-MSK) via topical administration for effective skin-whitening. To achieve this goal, EA and 4-MSK were formulated into lamellar liquid crystalline (LLC) cream, and response surface methodology (RSM) was employed to optimize the formulation. Polarized light microscopy (PLM), differential scanning calorimetry (DSC), and rheological experiments were performed to confirm the presence of the LLC structure in the base of cream. In addition, a comparison analysis of the skin retention of the two drugs between the LLC cream and the common o/w (COW) cream was made through in vitro permeation and in vivo drug distribution experiments. As a result, the optimal formulation was defined as 1.2% of EA, 1.48% of 4-MSK, 14.05% of Schercemol™ DISM Ester (DISM) as the oil, 4.0% of Emulium® Delta as the emulsifier, and 3.0% of stearyl alcohol as the co-emulsifier. In comparison with the COW cream, the LLC cream significantly increased the skin retention of EA and 4-MSK both in vitro and in vivo. In conclusion, the LLC carrier serves as a promising choice for topical preparation by enhancing skin retention and providing desirable rheological characteristics.

  14. Solar ultraviolet radiation induces biological alterations in human skin in vitro: Relevance of a well-balanced UVA/UVB protection

    Directory of Open Access Journals (Sweden)

    Françoise Bernerd

    2012-01-01

    Full Text Available Cutaneous damages such as sunburn, pigmentation, and photoaging are known to be induced by acute as well as repetitive sun exposure. Not only for basic research, but also for the design of the most efficient photoprotection, it is crucial to understand and identify the early biological events occurring after ultraviolet (UV exposure. Reconstructed human skin models provide excellent and reliable in vitro tools to study the UV-induced alterations of the different skin cell types, keratinocytes, fibroblasts, and melanocytes in a dose- and time-dependent manner. Using different in vitro human skin models, the effects of UV light (UVB and UVA were investigated. UVB-induced damages are essentially epidermal, with the typical sunburn cells and DNA lesions, whereas UVA radiation-induced damages are mostly located within the dermal compartment. Pigmentation can also be obtained after solar simulated radiation exposure of pigmented reconstructed skin model. Those models are also highly adequate to assess the potential of sunscreens to protect the skin from UV-associated damage, sunburn reaction, photoaging, and pigmentation. The results showed that an effective photoprotection is provided by broad-spectrum sunscreens with a potent absorption in both UVB and UVA ranges.

  15. Assessing the photoprotective effects of red ochre on human skin by in vitro laboratory experiments

    Directory of Open Access Journals (Sweden)

    Riaan F. Rifkin

    2015-03-01

    Full Text Available Archaeological indicators of cognitive complexity become increasingly prevalent during the African Middle Stone Age, with the habitual exploitation of red ochre widely viewed as a key feature of the emergence of modern human behaviour. Given that some of the uses of ochre remain ambiguous, we present the preliminary results of an ongoing study in which we explore the efficacy of red ochre as a photoprotective device or sunscreen. The capacity of ochre to inhibit the susceptibility of humans to the detrimental effects of ultraviolet radiation was confirmed through the in vitro calculation of the sun protection factor values of samples derived from the Kunene Region in Namibia and the Bokkeveld Group deposits, Western Cape Province, South Africa. Visible spectroscopy was employed to determine colourimetric parameters of samples and assess the correlation between ochre colour and sun protection factor. The possible role of ochre as a sunscreen agent for hominin populations, including modern humans, during the Middle Stone Age in Africa is explored. We conclude that the habitual use of red ochre as a photoprotective agent likely played a role in the ability of prehistoric humans to adapt to novel environmental circumstances.

  16. A genomic biomarker signature can predict skin sensitizers using a cell-based in vitro alternative to animal tests

    Directory of Open Access Journals (Sweden)

    Albrekt Ann-Sofie

    2011-08-01

    Full Text Available Abstract Background Allergic contact dermatitis is an inflammatory skin disease that affects a significant proportion of the population. This disease is caused by an adverse immune response towards chemical haptens, and leads to a substantial economic burden for society. Current test of sensitizing chemicals rely on animal experimentation. New legislations on the registration and use of chemicals within pharmaceutical and cosmetic industries have stimulated significant research efforts to develop alternative, human cell-based assays for the prediction of sensitization. The aim is to replace animal experiments with in vitro tests displaying a higher predictive power. Results We have developed a novel cell-based assay for the prediction of sensitizing chemicals. By analyzing the transcriptome of the human cell line MUTZ-3 after 24 h stimulation, using 20 different sensitizing chemicals, 20 non-sensitizing chemicals and vehicle controls, we have identified a biomarker signature of 200 genes with potent discriminatory ability. Using a Support Vector Machine for supervised classification, the prediction performance of the assay revealed an area under the ROC curve of 0.98. In addition, categorizing the chemicals according to the LLNA assay, this gene signature could also predict sensitizing potency. The identified markers are involved in biological pathways with immunological relevant functions, which can shed light on the process of human sensitization. Conclusions A gene signature predicting sensitization, using a human cell line in vitro, has been identified. This simple and robust cell-based assay has the potential to completely replace or drastically reduce the utilization of test systems based on experimental animals. Being based on human biology, the assay is proposed to be more accurate for predicting sensitization in humans, than the traditional animal-based tests.

  17. Mechanism of CW Nd:YAG laser recanalization with modified fiber tips: influence of temperature and axial force on tissue penetration in vitro.

    Science.gov (United States)

    Verdaasdonk, R M; Jansen, E D; Holstege, F C; Borst, C

    1991-01-01

    Modified fiber tips are used for laser angioplasty of totally occluded peripheral arteries. It has not been established, however, to what extent the mechanism of action of various laser probes is optical, thermal, or mechanical. We examined transparant contact probes (hemispherical contact probes and ball-shaped fibers) and metal laser probes, coupled to a continuous-wave Nd-YAG laser. By using homogeneous thick porcine fatty tissue samples submerged in blood plasma, tissue penetration was measured in relation to the temperature of the probe and the axial force exerted on the tissue. By using 15 W, 1 s laser pulses, the surface of transparent contact probes had to be first contaminated by carbonized tissue particles to achieve tissue penetration. Penetration increased from 1 to 10 mm per pulse when axial force increased from 20 to 100 g. Metal probes had to be sufficiently insulated from the liquid environment by water vapour entrapped in a denatured protein layer to exceed the threshold temperature of 225 degrees C for tissue penetration. When axial force increased from 20 to 80 g at 10 W continuous exposure, the velocity of tissue penetration increased in the range from 1 to 4 mm/s. Tissue penetration by modified fiber tips is attributed to both remodeling and vaporization of tissue. With transparent contact probes, tissue is heated partly by direct light absorption and partly by a hot probe surface. Axially directed force is necessary to displace lateral non-ablated tissue and to overcome mechanical resistance. We conclude that mechanical dilation due to axial catherization force (Dotter effect) contributes substantially to tissue penetration by transparent contact probes.

  18. Oligonucleotides suppress IL-8 in skin keratinocytes in vitro and offer anti-inflammatory properties in vivo.

    Science.gov (United States)

    Dorn, Annette; Ludwig, Ralf Joachim; Bock, Andreas; Thaci, Diamant; Hardt, Katja; Bereiter-Hahn, Jurgen; Kaufmann, Roland; Bernd, August; Kippenberger, Stefan

    2007-04-01

    DNA codes for genetic information. Furthermore, recent findings suggest that DNA offers additional function, particularly in the recognition of microorganisms. In this study, we investigated two classes of oligodeoxynucleotides (ODN) in skin keratinocytes; namely, an ODN comprising two cytidine-phosphate-guanosine (CpG) motifs (CpG-1-phosphorothioate (PTO)) and a poly-cytidine (Non-CpG-5-PTO) as control. Both fluorescence-tagged ODN were rapidly taken up by cells and accumulated already after 5 minutes in perinuclear compartments. In order to test whether ODN convey immunological effects in keratinocytes, secretion of IL-8 was measured. Interestingly, both CpG-1-PTO and Non-CpG-5-PTO suppressed basal and tumor necrosis factor alpha-induced IL-8 levels measured in cell culture supernatants. Experiments using deletion mutant revealed a critical length of approximately 16 nucleotides conveying IL-8 suppression. Studies regarding the ODN backbone offered that PTO bondings are critical for significant IL-8 suppression. In order to substantiate the anti-inflammatory response, a contact hypersensitivity mouse model was utilized. Topical application of Non-CpG-5-PTO-containing ointments reduced ear thickness in sensitized mice. Taken together, these findings suggest an anti-inflammatory effect of ODN in epithelial cells in vitro and in vivo, indicating that DNA molecules offer distinct biological activities restricted to the physiological compartment applied. This effect seems to be independent from Toll-like receptor 9.

  19. Skin-Derived Precursor Cells as an In Vitro Modelling Tool for the Study of Type 1 Neurofibromatosis

    Directory of Open Access Journals (Sweden)

    Araika Gutiérrez-Rivera

    2012-01-01

    Full Text Available The most characteristic feature of neurofibromatosis type 1 (NF1 is the development of neurofibromas. It has been suggested that these tumors are caused by somatic inactivation of the wild-type NF1 allele, but the cell that originally suffers this mutation remains controversial. Several lines of evidence support the clonal origin of these tumors, and it has been recently suggested that skin-derived precursor cells (SKPs could be the cell of origin of dermal neurofibromas. Nullizygous (NF1−/− SKPs do give rise to neurofibromas when transplanted to heterozygous mice. Moreover, a nullizygous population of cells that is S100β negative is present in human neurofibromas, and NF1+/− multipotent progenitor cells are seemingly recruited to the tumor. This evidence supports the neurofibroma stem cell hypothesis and a putative involvement of SKPs in the aetiopathogenesis of the disease, suggesting that SKPs could become a valuable tool for the in vitro study of NF1.

  20. Effects of industrial detergents on the barrier function of human skin

    DEFF Research Database (Denmark)

    Nielsen, G D; Nielsen, Jesper Bo; Andersen, Klaus Ejner

    2000-01-01

    and nickel was examined in an in-vitro model using human skin. Twenty-four of the most widely used detergents were studied. After a two-hour exposure to an aqueous detergent solution, penetration of labeled model compounds was followed for 66 hours. Interindividual variation was substantial, but 12...... of the detergents caused statistically significant increases in the penetration of water, nickel, or both. Nonionic detergents were as likely as anionic detergents to have this effect. This study demonstrates that useful information may be obtained by a simple in-vitro method, and that such data may provide a basis...

  1. Evaluation of EpiDerm full thickness-300 (EFT-300) as an in vitro model for skin irritation: Studies on aliphatic hydrocarbons

    Science.gov (United States)

    Mallampati, Ramya; Patlolla, Ram R.; Agarwal, Saurab; Babu, R. Jayachandra; Hayden, Patrick; Klausner, Mitchell; Singh, Mandip S.

    2010-01-01

    The aim of this study was to understand the skin irritation effects of saturated aliphatic hydrocarbons (HCs), C9–C16, found jet fuels using in vitro 3-dimensional EpiDerm full thickness-300 (EFT-300) skin cultures. The EFT-300 cultures were treated with 2.5 µl of HCs and the culture medium and skin samples were collected at 24 and 48 h to measure the release of various inflammatory biomarkers (IL-1α, IL-6 and IL-8). To validate the in vitro results, in vivo skin irritation studies were carried out in hairless rats by measuring trans epidermal water loss (TEWL) and erythema following un-occlusive dermal exposure of HCs for 72 h. The MTT tissue viability assay results with the EFT-300 tissue show that 2.5 µl/tissue (≈4.1 µl/cm2) of the HCs did not induce any significant changes in the tissue viability for exposure times up to 48 h of exposure. Microscopic observation of the EFT-300 cross-sections indicated that there were no obvious changes in the tissue morphology of the samples at 24 h, but after 48 h of exposure, tridecane, tetradecane and hexadecane produced a slight thickening and disruption of stratum corneum. Dermal exposures of C12–C16 HCs for 24 h significantly increased the expression of IL-1α in the skin as well as in the culture medium. Similarly, dermal exposure of all HCs for 24 h significantly increased the expression of interleukin-6 (IL-6) and IL-8 in the skin as well as in the culture medium in proportion to the HC chain length. As the exposure time increased to 48 h, IL-6 concentrations increased 2-fold compared to the IL-6 values at 24 h. The in vivo skin irritation data also showed that both TEWL and erythema scores increased with increased HCs chain length (C9–C16). In conclusion, the EFT-300 showed that the skin irritation profile of HCs was in the order of C9 ≤ C10 ≤ C11 ≤ C12 < C13 ≈ C14 ≈ C16 and that the tissue was an excellent in vitro model to predict in vivo irritation and to understand the structural activity

  2. Effect of postmortem time interval on in vitro culture potential of goat skin tissues stored at room temperature.

    Science.gov (United States)

    Singh, Mahipal; Ma, Xiaoling; Sharma, Anil

    2012-09-01

    Animal cloning using somatic cell nuclear transfer technology has renewed the interest in postmortem tissue storage, since these tissues can be used to reintroduce the lost genes back into the breeding pool in animal agriculture, preserve the genetic diversity, and revive the endangered species. However, for successful cloning of animals, integrity of nuclear DNA is essential. Cell viability and their potential to in vitro culture ensure nuclear integrity. The aim of this study was to determine the limits of postmortem time interval within which live cells can be recovered from goat skin tissues. To test the postmortem tissue storage limits, we cultured 2-3 mm(2) skin pieces (n = 70) from the ears of three breeds of goats (n = 7) after 0, 2, 4, and 6 days of postmortem storage at 24°C. After 10 days of culture, outgrowth of fibroblast-like cells (>50 cells) around the explants was scored. All the explants irrespective of breed displayed outgrowth of cells on the dish containing fresh tissues (i.e., day 0 of storage). However, the number of explants exhibiting outgrowth reduced with increasing time interval. Only 53.85 % explants displayed outgrowth after 2 days of tissue storage. The number of explants displaying outgrowth was much smaller after 4 (16.67 %) and 6 days (13.3 %) of storage. In general, the number of outgrowing cells per explant, on a given day, also decreased with increasing postmortem storage time interval. To test the differences between cell cultures, we established secondary cultures from one of the goats exhibiting outgrowth of cells after 6 days of tissue storage and compared them to similar cells from fresh tissues. Comparison of both the cell lines revealed similar cell morphology and growth curves and had doubling times of 23.04 and 22.56 h, respectively. These results suggest that live cells can be recovered from goat (and perhaps other animal) tissues stored at room temperature even after 6 days of their death with comparable growth

  3. The potential role of polyphenols in the modulation of skin cell viability by Aspalathus linearis and Cyclopia spp. herbal tea extracts in vitro.

    Science.gov (United States)

    Magcwebeba, Tandeka Unathi; Riedel, Sylvia; Swanevelder, Sonja; Swart, Pieter; De Beer, Dalene; Joubert, Elizabeth; Andreas Gelderblom, Wentzel Christoffel

    2016-11-01

    The relationship between polyphenol constituents, antioxidant properties of aqueous and methanol extracts of green tea (Camellia sinensis), the herbal teas, rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.), against skin cell viability was investigated in vitro. The effect of extracts, characterised in terms of polyphenol content and antioxidant properties, on cell viability of premalignant, normal and malignant skin cells was determined. Phenolic composition, particularly high levels of potent antioxidants, of rooibos and green tea methanol extracts was associated with a strong reduction in cell viability specifically targeting premalignant cells. In contrast, the aqueous extracts of Cyclopia spp. were more effective in reducing cell viability. This correlated with a relatively high flavanol/proanthocyanidin content and ABTS radical cation scavenging capacity. The major green tea flavanol (epigallocatechin gallate) and rooibos dihydrochalcone (aspalathin) exhibited differential effects against cell viability, while the major honeybush xanthone (mangiferin) and flavanone (hesperidin) lacked any effect presumably due to a cytoprotective effect. The underlying mechanisms against skin cell viability are likely to involve mitochondrial dysfunction resulting from polyphenol-iron interactions. The polyphenol constituents and antioxidant parameters of herbal tea extracts are useful tools to predict their activity against skin cell survival in vitro and potential chemopreventive effects in vivo. © 2016 Royal Pharmaceutical Society.

  4. Extracts from Calendula officinalis offer in vitro protection against H2 O2 induced oxidative stress cell killing of human skin cells.

    Science.gov (United States)

    Alnuqaydan, Abdullah M; Lenehan, Claire E; Hughes, Rachel R; Sanderson, Barbara J

    2015-01-01

    The in vitro safety and antioxidant potential of Calendula officinalis flower head extracts was investigated. The effect of different concentrations (0.125, 0.5, 1.0, 2.0 and 5.0% (v/v)) of Calendula extracts on human skin cells HaCaT in vitro was explored. Doses of 1.0% (v/v) (0.88 mg dry weight/mL) or less showed no toxicity. Cells were also exposed to the Calendula extracts for either 4, 24 or 48 h before being exposed to an oxidative insult (hydrogen peroxide H2 O2 ) for 1 h. Using the MTT cytotoxicity assay, it was observed that two independent extracts of C. officinalis gave time-dependent and concentration-dependent H2 O2 protection against induced oxidative stress in vitro using human skin cells. Pre-incubation with the Calendula extracts for 24 and 48 h increased survival relative to the population without extract by 20% and 40% respectively following oxidative challenge. The antioxidant potential of the Calendula extracts was confirmed using a complimentary chemical technique, the DPPH(●) assay. Calendula extracts exhibited free radical scavenging abilities. This study demonstrates that Calendula flower extracts contain bioactive and free radical scavenging compounds that significantly protect against oxidative stress in a human skin cell culture model. Copyright © 2014 John Wiley & Sons, Ltd.

  5. Chlorine and hydrogen cyanide gas interactions with human skin: in vitro studies to inform skin permeation and decontamination in HAZMAT incidents.

    Science.gov (United States)

    Gaskin, Sharyn; Pisaniello, Dino; Edwards, John W; Bromwich, David; Reed, Sue; Logan, Michael; Baxter, Christina

    2013-11-15

    Accidental or intentional toxic gas releases may result in significant public health and psychological consequences. Management of exposed individuals during HAZMAT incidents should be risk-based and supported by a suitable scientific evidence base. There appear to be large evidence gaps in relation to dermal absorption of gases, as well as management advice for potentially exposed individuals. Chlorine and hydrogen cyanide are two common HAZMAT gases and this paper addresses the need for experimental data tailored to HAZMAT scenarios and first responders. In addition to time variations of gas concentration, the modifying effects of clothing, temperature, and oil-based sunscreen on epidermal absorption and penetration are assessed. Results for chlorine show little penetration up to 500 ppm but with small enhancing effects due to heavy cotton and oil-based sunscreen. Hydrogen cyanide up to 800 ppm shows minor penetration consistent with previous studies, with little variability in the presence of sunscreen and clothing. Practical guidelines to support the decision-making of emergency responders with regard to personal decontamination have been derived. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. In-vitro permeability of the human nail and of a keratin membrane from bovine hooves: penetration of chloramphenicol from lipophilic vehicles and a nail lacquer.

    Science.gov (United States)

    Mertin, D; Lippold, B C

    1997-03-01

    Lipophilic vehicles and especially nail lacquers are more appropriate for topical application on the nail than aqueous systems because of their better adhesion. This work has, therefore, studied the penetration through the human nail plate of the model compound chloramphenicol from the lipophilic vehicles medium chain triglycerides and n-octanol and from a lacquer based on quaternary poly(methyl methacrylates) (Eudragit RL). The results were compared with data obtained with a keratin membrane from bovine hooves. If the swelling of the nail plate or the hoof membrane is not altered by use of lipophilic vehicles, the maximum flux of the drug is independent of its solubility in the vehicle and is the same as that from a saturated aqueous solution. These vehicles are not able to enter the hydrophilic keratin membrane because of their non-polar character and so cannot change the solubility of the penetrating substance in the barrier. If the concentration of the drug in the nail lacquer is sufficiently high, the maximum flux through both barriers equals that from aqueous vehicles or even exceeds it because of the formation of a supersaturated system. Penetration through the nail plate follows first order kinetics after a lag-time of 400 h. The course of penetration through the hoof membrane is initially membrane-controlled and later becomes a matrix-controlled process because of the membrane's greater permeability. Chloramphenicol is dissolved in the lacquer up to a concentration of 31%. The relative release rates from these solution matrices are independent of the drug concentration but they decrease on changing to a suspension matrix. These results show that drug flux is independent of the character of the vehicle and that penetration of the drug is initially membrane-controlled and changes to being matrix-controlled as the drug content of the lacquer decreases.

  7. Antibacterial Evaluation of Synthetic Thiazole Compounds In Vitro and In Vivo in a Methicillin-Resistant Staphylococcus aureus (MRSA) Skin Infection Mouse Model

    Science.gov (United States)

    Mohammad, Haroon; Cushman, Mark; Seleem, Mohamed N.

    2015-01-01

    The emergence of community-associated methicillin-resistant Staphylococcus aureus (MRSA), including strains resistant to current antibiotics, has contributed to an increase in the number of skin infections reported in humans in recent years. New therapeutic options are needed to counter this public health challenge. The aim of the present study was to examine the potential of thiazole compounds synthesized by our research group to be used topically to treat MRSA skin and wound infections. The broth microdilution method confirmed that the lead thiazole compound and four analogues are capable of inhibiting MRSA growth at concentrations as low as 1.3 μg/mL. Additionally, three compounds exhibited a synergistic relationship when combined with the topical antibiotic mupirocin against MRSA in vitro via the checkerboard assay. Thus the thiazole compounds have potential to be used alone or in combination with mupirocin against MRSA. When tested against human keratinocytes, four derivatives of the lead compound demonstrated an improved toxicity profile (were found to be non-toxic up to a concentration of 20 μg/mL). Utilizing a murine skin infection model, we confirmed that the lead compound and three analogues exhibited potent antimicrobial activity in vivo, with similar capability as the antibiotic mupirocin, as they reduced the burden of MRSA present in skin wounds by more than 90%. Taken altogether, the present study provides important evidence that these thiazole compounds warrant further investigation for development as novel topical antimicrobials to treat MRSA skin infections. PMID:26536129

  8. Deformable liposomes and ethosomes: mechanism of enhanced skin delivery.

    Science.gov (United States)

    Elsayed, Mustafa M A; Abdallah, Ossama Y; Naggar, Viviane F; Khalafallah, Nawal M

    2006-09-28

    Despite intensive research, the mechanisms by which vesicular systems deliver drugs into intact skin are not yet fully understood. In the current study, possible mechanisms by which deformable liposomes and ethosomes improve skin delivery of ketotifen under non-occlusive conditions were investigated. In vitro permeation and skin deposition behavior of deformable liposomes and ethosomes, having ketotifen both inside and outside the vesicles (no separation of free ketotifen), having ketotifen only inside the vesicles (free ketotifen separated) and having ketotifen only outside the vesicles (ketotifen solution added to empty vesicles), was studied using rabbit pinna skin. Results suggested that both the penetration enhancing effect and the intact vesicle permeation into the stratum corneum might play a role in improving skin delivery of drugs by deformable liposomes, under non-occlusive conditions, and that the penetration enhancing effect was of greater importance in case of ketotifen. Regarding ethosomes, results indicated that ketotifen should be incorporated in ethosomal vesicles for optimum skin delivery. Ethosomes were not able to improve skin delivery of non-entrapped ketotifen.

  9. A new dermocosmetic containing retinaldehyde, delta-tocopherol glucoside and glycylglycine oleamide for managing naturally aged skin: results from in vitro to clinical studies

    Science.gov (United States)

    Rouvrais, Céline; Bacqueville, Daniel; Bogdanowicz, Patrick; Haure, Marie-José; Duprat, Laure; Coutanceau, Christine; Castex-Rizzi, Nathalie; Duplan, Hélène; Mengeaud, Valérie; Bessou-Touya, Sandrine

    2017-01-01

    Introduction Natural aging of skin tissues, the addition of the cumulative action of the time and radiation exposure result in skin atrophy, wrinkles and degeneration of the extracellular matrix (ECM). The aim of the study was to investigate the beneficial effect of a combination containing retinaldehyde (RAL), delta-tocopherol glucoside (delta-TC) and glycylglycine ole-amide (GGO) and of a dermocosmetic containing the combination. Materials and methods The protective effect of the combination was assessed through in vitro gene expression of ultraviolet (UV)-irradiated fibroblasts. A skin aging assay using UV light on ex vivo skin samples and a clinical study conducted in 36 women aged from 35 to 55 years with a minimum of level 4 to a maximum of level 6 on the crow’s feet photoscale assessed the antiaging effect of the dermocosmetic. Results When added to UV-irradiated fibroblasts, the combination substantially improved the ECM in activating the elastin fiber production (fibrillin 2, fibulin 1 and 5 and lysyl oxidase-like 2) as well as that of proteins involved in the cellular ECM interactions (integrin b1, paxillin and actin a2). An ex vivo photodamaged human skin model showed that the dermocosmetic formulation containing the combination of the active ingredients protected the elastic network against UV-induced alterations including both elastin and fibrillin-rich fibers in the dermis. A daily application of the dermocosmetic for 2 months on naturally aged skin resulted in a statistically significant improvement (pwrinkles and periocular fine lines. Finally, the formulation was well tolerated. Conclusion The dermocosmetic containing RAL, delta-TC and GGO provides a substantial benefit in the daily care of naturally aged skin in women aged 35–55 years. PMID:28203099

  10. Assessment of the percutaneous penetration of indomethacin from soybean oil microemulsion: effects of the HLB value of mixed surfactants.

    Science.gov (United States)

    Chen, Liangmei; Tan, Fengping; Wang, Jinfeng; Liu, Feng

    2012-01-01

    The objective of this study was to evaluate the influence of the ratios or the hydrophile-lipophile balance (HLB) values of Cremophor EL and Span 80 on the phase behavior of the O/W microemulsions and the percutaneous absorption and penetration of indomethacin microemulsions. The existence of microemulsion regions is investigated in quaternary systems composed of soybean oil/Cremophor EL and Span 80 (mixed surfactants)/diethylene glycol monoethyl ether (cosurfactant)/water by constructing pseudo-ternary phase diagrams at various Cremophor EL/Span 80 ratios. In addition, five microemulsion formulations with various mixed surfactants HLB values were evaluated by in vitro penetration experiments using mouse skin and Franz diffusion cells. The flux and amount of indomethacin penetration from 5 microemulsion formulations were significantly different from the control, and the enhance ratios ranged from 2.38 to 4.68 and 2.11 to 4.23, respectively. The HLB value of mixed surfactants in the formulations was a principal factor in determining the percutaneous penetration of the drug. The flux and amount of drug penetration increased gradually with increasing content of the lipophilic surfactant Span 80 and skin retention was highest for mixed surfactants with a HLB value of 7.6. Therefore, it is suggested that the presence of mixed surfactants was beneficial in the formation of O/W microemulsions and enhanced percutaneous penetration of indomethacin.

  11. Effect of technique of sealer agitation on percentage and depth of MTA Fillapex sealer penetration: A comparative in-vitro study

    Science.gov (United States)

    Nikhil, Vineeta; Bansal, Parul; Sawani, Shefali

    2015-01-01

    Aim: To compare the effect of three root canal sealer activation techniques on percentage and depth of sealer penetration of MTA Fillapex and AH Plus sealers. Materials and Methods: Sixty teeth prepared till F5 ProTaper size were divided into three equal groups on the basis of sealer activation technique (G1: Ultrasonics, G2: Lentulo spiral, and G3: Counter-clockwise rotary motion). Each group was further divided into two equal subgroups on the basis of type of sealer used: AH Plus (Denstply, Konstanz, Germany) or MTA Fillapex (Angelus, Londrina, PR, Brazil) and obturated with gutta-percha. Horizontal sections at 3 and 6 mm from the apex were obtained and the percentage and depth of penetration of sealers into dentinal tubules were measured using confocal laser scanning microscopy (CLSM). Statistical analysis was performed utilizing Kruskal-Wallis and Mann-Whitney U tests with a significance level of 5%. Results: G1 showed significantly (P 0.05) between G2 and G3. Conclusion: Percentage and depth of sealer penetration are influenced by the type of sealer used sealer activation technique and by the root canal level. Ultrasonic method of sealer activation and MTA Fillapex showed the best results. PMID:25829689

  12. Xenobiotic metabolism capacities of human skin in comparison with a 3D-epidermis model and keratinocyte-based cell culture as in vitro alternatives for chemical testing: phase II enzymes.

    Science.gov (United States)

    Götz, Christine; Pfeiffer, Roland; Tigges, Julia; Ruwiedel, Karsten; Hübenthal, Ulrike; Merk, Hans F; Krutmann, Jean; Edwards, Robert J; Abel, Josef; Pease, Camilla; Goebel, Carsten; Hewitt, Nicola; Fritsche, Ellen

    2012-05-01

    The 7th Amendment to the EU Cosmetics Directive prohibits the use of animals in cosmetic testing for certain endpoints, such as genotoxicity. Therefore, skin in vitro models have to replace chemical testing in vivo. However, the metabolic competence neither of human skin nor of alternative in vitro models has so far been fully characterized, although skin is the first-pass organ for accidentally or purposely (cosmetics and pharmaceuticals) applied chemicals. Thus, there is an urgent need to understand the xenobiotic-metabolizing capacities of human skin and to compare these activities to models developed to replace animal testing. We have measured the activity of the phase II enzymes glutathione S-transferase, UDP-glucuronosyltransferase and N-acetyltransferase in ex vivo human skin, the 3D epidermal model EpiDerm 200 (EPI-200), immortalized keratinocyte-based cell lines (HaCaT and NCTC 2544) and primary normal human epidermal keratinocytes. We show that all three phase II enzymes are present and highly active in skin as compared to phase I. Human skin, therefore, represents a more detoxifying than activating organ. This work systematically compares the activities of three important phase II enzymes in four different in vitro models directly to human skin. We conclude from our studies that 3D epidermal models, like the EPI-200 employed here, are superior over monolayer cultures in mimicking human skin xenobiotic metabolism and thus better suited for dermatotoxicity testing. © 2012 John Wiley & Sons A/S.

  13. The in vitro skin irritation of chemicals: optimisation of the EPISKIN prediction model within the framework of the ECVAM validation process.

    Science.gov (United States)

    Cotovio, Jose; Grandidier, Marie-Helene; Portes, Pascal; Roguet, Roland; Rubinstenn, Gilles

    2005-08-01

    In view of the increasing need to identify non-animal tests able to predict acute skin irritation of chemicals, the European Centre for the Validation of Alternative Methods (ECVAM) focused on the evaluation of appropriate in vitro models. In vitro tests should be capable of discriminating between irritant (I) chemicals (EU risk: R38) and non-irritant (NI) chemicals (EU risk: "no classification"). Since major in vivo skin irritation assays rely on visual scoring, it is still a challenge to correlate in vivo clinical signs with in vitro biochemical measurements. Being particularly suited to test raw materials or chemicals with a wide variety of physical properties, in vitro skin models resembling in vivo human skin were involved in prevalidation processes. Among many other factors, cytotoxicity is known to trigger irritation processes, and can therefore be a first common event for irritants. A refined protocol (protocol 15min-18hours) for the EPISKIN model had been proposed for inclusion in the ECVAM formal validation study. A further improvement on this protocol, mainly based on a post-treatment incubation period of 42 hours (protocol 15min-42hours), the optimised protocol, was applied to a set of 48 chemicals. The sensitivity, specificity and accuracy with the MTT assay-based prediction model (PM) were 85%, 78.6% and 81.3% respectively, with a low rate of false negatives (12%). The improved performance of this optimised protocol was confirmed by a higher robustness (homogeneity of individual responses) and a better discrimination between the I and NI classes. To improve the MTT viability-based PM, the release of a membrane damage marker, adenylate kinase (AK), and of cytokines IL-1alpha and IL-8 were also investigated. Combining these endpoints, a simple two-tiered strategy (TTS) was developed, with the MTT assay as the first, sort-out, stage. This resulted in a clear increase in sensitivity to 95%, and a fall in the false-positive rate (to 4.3%), thus

  14. Effects of single and repeated exposure to biocidal active substances on the barrier function of the skin in vitro

    NARCIS (Netherlands)

    Buist, H.E.; Sandt, J.J.M. van de; Burgsteden, J.A. van; Heer, C. de

    2005-01-01

    The dermal route of exposure is important in worker exposure to biocidal products. Many biocidal active substances which are used on a daily basis may decrease the barrier function of the skin to a larger extent than current risk assessment practice addresses, due to possible skin effects of

  15. ZEB1 is Estrogen Responsive In Vitro in Human Foreskin Cells and is Over Expressed in Penile Skin in Patients With Severe Hypospadias

    Science.gov (United States)

    Qiao, Liang; Tasian, Gregory E.; Zhang, Haiyang; Cunha, Gerald R.; Baskin, Laurence

    2012-01-01

    Purpose We determined the effect of estrogen on ZEB1 in vitro and tested the hypothesis that ZEB1 is over expressed in the penile skin of subjects with hypospadias. Materials and Methods Hs68 cells, a fibroblast cell line derived from human foreskin, were exposed to 0, 1, 10 and 100 nM estrogen, and the expression level of ZEB1 was assessed using reverse transcription real-time polymerase chain reaction, Western blot and immunocytochemical analysis. Next, preputial skin was prospectively collected from case and control subjects at hypospadias repair (37 cases) and circumcision (11). Hypospadias was classified as severe (13 cases) or mild (24) based on the position of the urethral meatus. ZEB1 expression was quantified using reverse transcription real-time polymerase chain reaction, Western blot and immunohistochemical analysis. Results Estrogen increased ZEB1 expression at the mRNA and protein levels in Hs68 cells in a concentration dependent fashion (p hypospadias had significantly higher ZEB1 mRNA levels and protein expression compared to controls or subjects with mild hypospadias (both p hypospadias had increased expression of ZEB1 in the basal layers of the preputial epidermis. Conclusions Estrogen increases ZEB1 expression in a human foreskin fibroblast cell line in vitro. Furthermore, ZEB1 is significantly over expressed in the penile skin of subjects with severe hypospadias. We propose that ZEB1 overexpression may contribute to development of hypospadias and may mediate the effect of estrogen on developing external male genitalia. PMID:21421232

  16. In vitro-in vivo correlations for nicotine transdermal delivery systems evaluated by both in vitro skin permeation (IVPT) and in vivo serum pharmacokinetics under the influence of transient heat application.

    Science.gov (United States)

    Shin, Soo Hyeon; Thomas, Sherin; Raney, Sam G; Ghosh, Priyanka; Hammell, Dana C; El-Kamary, Samer S; Chen, Wilbur H; Billington, M Melissa; Hassan, Hazem E; Stinchcomb, Audra L

    2018-01-28

    The in vitro permeation test (IVPT) has been widely used to characterize the bioavailability (BA) of compounds applied on the skin. In this study, we performed IVPT studies using excised human skin (in vitro) and harmonized in vivo human serum pharmacokinetic (PK) studies to evaluate the potential in vitro-in vivo correlation (IVIVC) of nicotine BA from two, matrix-type, nicotine transdermal delivery systems (TDS). The study designs used for both in vitro and in vivo studies included 1h of transient heat (42±2°C) application during early or late time periods post-dosing. The goal was to evaluate whether any IVIVC observed would be evident even under conditions of heat exposure, in order to investigate further whether IVPT may have the potential to serve as a possible surrogate method to evaluate the in vivo effects of heat on the bioavailability of a drug delivered from a TDS. The study results have demonstrated that the BA of nicotine characterized by the IVPT studies correlated with and was predictive of the in vivo BA of nicotine from the respective TDS, evaluated under the matched study designs and conditions. The comparisons of single parameters such as steady-state concentration, heat-induced increase in partial AUCs and post-treatment residual content of nicotine in TDS from the in vitro and in vivo data sets showed no significant differences (p≥0.05). In addition, a good point-to-point IVIVC (Level A correlation) for the entire study duration was achieved by predicting in vivo concentrations of nicotine using two approaches: Approach I requiring only an in vitro data set and Approach II involving deconvolution and convolution steps. The results of our work suggest that a well designed IVPT study with adequate controls can be a useful tool to evaluate the relative effects of heat on the BA of nicotine from TDS with different formulations. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Sacha Inchi Oil (Plukenetia volubilis L.), effect on adherence of Staphylococus aureus to human skin explant and keratinocytes in vitro.

    Science.gov (United States)

    Gonzalez-Aspajo, German; Belkhelfa, Haouaria; Haddioui-Hbabi, Laïla; Bourdy, Geneviève; Deharo, Eric

    2015-08-02

    Plukenetia volubilis L. (Euphorbiaceae) is a domesticated vine distributed from the high-altitude Andean rain forest to the lowlands of the Peruvian Amazon. Oil from the cold-pressed seeds, sold under the commercial name of Sacha Inchi Oil (SIO) is actually much in favour because it contains a high percentage of omega 3 and omega 6, and is hence used as a dietary supplement. SIO is also used traditionally for skin care, in order to maintain skin softness, and for the treatment of wounds, insect bites and skin infections, in a tropical context where the skin is frequently damaged. This study was designed in order to verify whether the traditional use of SIO for skin care would have any impact on Staphylococcus aureus growth and skin adherence, as S. aureus is involved in many skin pathologies (impetigo, folliculitis, furuncles and subcutaneous abscesses) being one if the main pathogens that can be found on the skin. Therefore, our objective was to assess SIO bactericidal activity and interference with adherence to human skin explants and the keratinocyte cell line. Cytotoxicity on that cells was also determined. The activity of SIO was compared to coconut oil (CocO), which is widely used for skin care but has different unsaturated fatty acids contents. Laboratory testing with certified oil, determined antibacterial activity against radio labelled S. aureus. Cytotoxic effects were measured with XTT on keratinocyte cells and with neutral red on human skin explants; phenol was used as cytotoxic control. Adherence assays were carried out by mixing H3-labelled S. aureus bacteria with keratinocyte cells and human skin explants, incubated with oils 2h before (to determine the inhibition of adherence, assimilated to a preventive effect) or 2h after the contact of the biological material with S. aureus (to assess the detachment of the bacteria, assimilated to a curative effect). Residual radioactivity measured after washings made it possible to determine the adherence

  18. Skin barrier function

    DEFF Research Database (Denmark)

    2016-01-01

    on the subject. It covers new basic research on skin markers, including results on filaggrin and on methods for the assessment of the barrier function. Biological variation and aspects of skin barrier function restoration are discussed as well. Further sections are dedicated to clinical implications of skin...... barrier integrity, factors influencing the penetration of the skin, influence of wet work, and guidance for prevention and saving the barrier. Distinguished researchers have contributed to this book, providing a comprehensive and thorough overview of the skin barrier function. Researchers in the field...

  19. Improvement of skin optical clearing efficacy by topical treatment of glycerol at different temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Deng Zijian; Liu Caihua; Tao Wei; Zhu Dan, E-mail: dawnzh@mail.hust.edu.cn [Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan (China)

    2011-01-01

    In the past decades, laser has been widely used in clinical diagnosis and cosmetic therapy. However, there is limitation for further usage in deeper tissue for high scattering property. Skin optical clearing technique, by introducing optical clearing agents (OCAs) into tissue, will have a potential impact on optical diagnosis and therapy. In this work, anhydrous glycerol at different temperatures of 4, 25, 32 and 45 deg. C were applied respectively to in vitro porcine skin, and reflectance and transmittance spectra were then measured dynamically using a spectrometry combined with integrating sphere system. Further, reduced scattering coefficient and penetration depth were obtained. Results showed that, glycerol at different temperatures could induce the reduced scattering coefficient of in vitro skin to decrease and the penetration depth to increase. 4 and 25 deg. C glycerol had similar effect, decreasing the scattering by 48.2% and 49.7%, and increasing penetration depth by 37.9% and 39.5%, respectively. However, 32 and 45 deg. C glycerol treatment could decrease scattering by 61.6% and 76.6%, and increase penetration depth by 53.3% and 84.1%, respectively. In conclusion, glycerol at higher temperature can induce greater and faster skin optical clearing efficacy.

  20. Improvement of skin optical clearing efficacy by topical treatment of glycerol at different temperatures

    Science.gov (United States)

    Deng, Zijian; Liu, Caihua; Tao, Wei; Zhu, Dan

    2011-01-01

    In the past decades, laser has been widely used in clinical diagnosis and cosmetic therapy. However, there is limitation for further usage in deeper tissue for high scattering property. Skin optical clearing technique, by introducing optical clearing agents (OCAs) into tissue, will have a potential impact on optical diagnosis and therapy. In this work, anhydrous glycerol at different temperatures of 4, 25, 32 and 45°C were applied respectively to in vitro porcine skin, and reflectance and transmittance spectra were then measured dynamically using a spectrometry combined with integrating sphere system. Further, reduced scattering coefficient and penetration depth were obtained. Results showed that, glycerol at different temperatures could induce the reduced scattering coefficient of in vitro skin to decrease and the penetration depth to increase. 4 and 25°C glycerol had similar effect, decreasing the scattering by 48.2% and 49.7%, and increasing penetration depth by 37.9% and 39.5%, respectively. However, 32 and 45°C glycerol treatment could decrease scattering by 61.6% and 76.6%, and increase penetration depth by 53.3% and 84.1%, respectively. In conclusion, glycerol at higher temperature can induce greater and faster skin optical clearing efficacy.

  1. A new dermocosmetic containing retinaldehyde, delta-tocopherol glucoside and glycylglycine oleamide for managing naturally aged skin: results from in vitro to clinical studies

    Directory of Open Access Journals (Sweden)

    Rouvrais C

    2017-02-01

    Full Text Available Céline Rouvrais,1,* Daniel Bacqueville,2,* Patrick Bogdanowicz,2,* Marie-José Haure,2 Laure Duprat,2 Christine Coutanceau,3 Nathalie Castex-Rizzi,2 Hélène Duplan,2 Valérie Mengeaud,1 Sandrine Bessou-Touya2 1Clinical Skin Research Center, 2Department of Pharmacology, Pierre Fabre Dermo-Cosmétique, Toulouse, 3Laboratoire Dermatologique Avène, Lavaur, France *These authors contributed equally to this work Introduction: Natural aging of skin tissues, the addition of the cumulative action of the time and radiation exposure result in skin atrophy, wrinkles and degeneration of the extracellular matrix (ECM. The aim of the study was to investigate the beneficial effect of a combination containing retinaldehyde (RAL, delta-tocopherol glucoside (delta-TC and glycylglycine oleamide (GGO and of a dermocosmetic containing the combination. Materials and methods: The protective effect of the combination was assessed through in vitro gene expression of ultraviolet (UV-irradiated fibroblasts. A skin aging assay using UV light on ex vivo skin samples and a clinical study conducted in 36 women aged from 35 to 55 years with a minimum of level 4 to a maximum of level 6 on the crow’s feet photoscale assessed the antiaging effect of the dermocosmetic. Results: When added to UV-irradiated fibroblasts, the combination substantially improved the ECM in activating the elastin fiber production (fibrillin 2, fibulin 1 and 5 and lysyl ­oxidase-like 2 as well as that of proteins involved in the cellular ECM interactions (integrin β1, paxillin and actin a2. An ex vivo photodamaged human skin model showed that the dermocosmetic formulation containing the combination of the active ingredients protected the elastic network against UV-induced alterations including both elastin and fibrillin-rich fibers in the dermis. A daily application of the dermocosmetic for 2 months on naturally aged skin resulted in a statistically significant improvement (p<0.05 of visible

  2. Germ tube mediated invasion of Batrachochytrium dendrobatidis in amphibian skin is host dependent.

    Directory of Open Access Journals (Sweden)

    Pascale Van Rooij

    Full Text Available Batrachochytrium dendrobatidis (Bd is the causative agent of chytridiomycosis, a fungal skin disease in amphibians and driver of worldwide amphibian declines.We focussed on the early stages of infection by Bd in 3 amphibian species with a differential susceptibility to chytridiomycosis. Skin explants of Alytes muletensis, Litoria caerulea and Xenopus leavis were exposed to Bd in an Ussing chamber for 3 to 5 days. Early interactions of Bd with amphibian skin were observed using light microscopy and transmission electron microscopy. To validate the observations in vitro, comparison was made with skin from experimentally infected frogs. Additional in vitro experiments were performed to elucidate the process of intracellular colonization in L. caerulea. Early interactions of Bd with amphibian skin are: attachment of zoospores to host skin, zoospore germination, germ tube development, penetration into skin cells, invasive growth in the host skin, resulting in the loss of host cell cytoplasm. Inoculation of A. muletensis and L. caerulea skin was followed within 24 h by endobiotic development, with sporangia located intracellularly in the skin. Evidence is provided of how intracellular colonization is established and how colonization by Bd proceeds to deeper skin layers. Older thalli develop rhizoid-like structures that spread to deeper skin layers, form a swelling inside the host cell to finally give rise to a new thallus. In X. laevis, interaction of Bd with skin was limited to an epibiotic state, with sporangia developing upon the skin. Only the superficial epidermis was affected. Epidermal cells seemed to be used as a nutrient source without development of intracellular thalli. The in vitro data agreed with the results obtained after experimental infection of the studied frog species. These data suggest that the colonization strategy of B. dendrobatidis is host dependent, with the extent of colonization most likely determined by inherent

  3. Photoacoustic study of percutaneous absorption of Carbopol and transdermic gels for topic use in skin

    Science.gov (United States)

    Rossi, R. C. P.; de Paiva, R. F.; da Silva, M. D.; Barja, P. R.

    2008-01-01

    Topical medicine application has been used to treat a good number of pathological processes. Its efficacy is associated to an efficient penetration of the drug in the internal skin layers, promoting systemic effects and excluding the possibility of drug degradation by the digestive tract and hepatic elimination. This work analyzes the penetration kinetics of two soluble bases employed as vehicles for topic application: superficial gel (Carbopol 940) and transdermic (transdermal) gel. Analysis was performed with the photoacoustic technique, based upon the absorption of modulated light by a sample with subsequent conversion of the absorbed energy in heat, generating acoustic waves in the air layer adjacent to the sample. Each of the two vehicles was evaluated through in vivo (human skin) and in vitro application. Measurements in vitro employed samples of VitroSkin (synthetic material with properties similar to those of real skin, employed in the pharmaceutical industry research). Results show that the permeation was faster for the transdermal gel, both for in vivo and in vitro measurements, indicating that in vitro measurements may be utilized in qualitative, comparative permeation studies.

  4. Skin absorption through atopic dermatitis skin

    DEFF Research Database (Denmark)

    Halling-Overgaard, A-S; Kezic, S; Jakasa, I

    2017-01-01

    Patients with atopic dermatitis have skin barrier impairment in both lesional and non-lesional skin. They are typically exposed to emollients daily and topical anti-inflammatory medicaments intermittently, hereby increasing the risk of developing contact allergy and systemic exposed to chemicals...... ingredients found in these topical preparations. We systematically searched for studies that investigated skin absorption of various penetrants, including medicaments, in atopic dermatitis patients, but also animals with experimentally induced dermatitis. We identified 40 articles, i.e. 11 human studies...... examining model penetrants, 26 human studies examining atopic dermatitis drugs and 3 animal studies. We conclude that atopic dermatitis patients have nearly two-fold increased skin absorption when compared to healthy controls. There is a need for well-designed epidemiological and dermato...

  5. Skin permeation of lidocaine from crystal suspended oily formulations.

    Science.gov (United States)

    Matsui, Rakan; Hasegawa, Masaaki; Ishida, Masami; Ebata, Toshiya; Namiki, Noriyuki; Sugibayashi, Kenji

    2005-09-01

    In vitro permeation of lidocaine (lidocaine base, LID) through excised rat skin was investigated using several LID-suspended oily formulations. The first skin permeation of LID from an LID-suspended oily solution such as liquid paraffin (LP), isopropyl myristate (IPM), polyoxyethylene (2) oleylether (BO-2), and diethyl sebacate (DES) was evaluated and compared with that from polyethylene glycol 400 (PEG400) solution, a hydrophilic base. The obtained permeation rate of LID, Japp, from PEG400, LP, IPM, BO-2, and DES was in the order of DES>BO-2=IPM>LP>PEG400, and increased with LID solubility in the oily solvents, although LID crystals were dispersed in all solvents. Subsequently, oily formulations that consisted of different ratios of the first oily solvent (IPM, BO-2, or DES) (each 0-20%), the second oily solvent (LP) and an oily mixture of microcrystalline wax/white petrolatum/paraffin (1/5/4) were evaluated. BO-2 groups at a concentration of 5% and 10% had the highest Japp among the oily formulations, although a higher BO-2 resulted in lower skin permeation. In addition, pretreatment with BO-2 increased the skin permeation of LID. These results suggest that the penetration enhancing effect by the system may be related to the skin penetration of BO-2 itself. Finally, mathematical analysis was done to evaluate the effect of BO-2, and it was shown that BO-2 improved the LID solubility in stratum corneum lipids to efficiently enhance the LID permeation through skin.

  6. Tissue-engineered human psoriatic skin supplemented with cytokines as an in vitro model to study plaque psoriasis.

    Science.gov (United States)

    Pouliot-Bérubé, Claudia; Zaniolo, Karine; Guérin, Sylvain L; Pouliot, Roxane

    2016-09-01

    Psoriasis is a chronic inflammatory skin disease. To study its complex etiology, a psoriatic skin substitute model supplemented with a cytokine cocktail has been used. Reconstructed psoriatic skin substitutes were supplemented with a cocktail of four cytokines: TNF-α, IL-1α, IL-6 and IL-17A, to monitor their impact on gene expression by DNA microarray. Gene profiling analyses identified several deregulated genes reported as being also deregulated in psoriasis skin in vivo (S100A12, IL-8, DEFB4A and KYNU). The expression of those genes was dramatically increased compared with basal levels of controls (p < 0.005 to < 0.05). Psoriatic substitutes supplemented with a cocktail of TNF-α, IL-1α, IL-6 and IL-17A showed similar transcriptome alterations to those found in psoriasis.

  7. Physiochemical properties and resorption progress of porcine skin-derived collagen membranes: In vitro and in vivo analysis

    National Research Council Canada - National Science Library

    AN, Yin-Zhe; KIM, You-Kyoung; LIM, Su-Min; HEO, Yeong-Ku; KWON, Mi-Kyung; CHA, Jae-Kook; LEE, Jung-Seok; JUNG, Ui-Won; CHOI, Seong-Ho

    2018-01-01

    The aim of the present study was to evaluate the physiochemical properties and resorption progress of two cross-linked, porcine skin-derived collagen membranes and compare their features with those of...

  8. Listening to mozart reduces allergic skin wheal responses and in vitro allergen-specific IgE production in atopic dermatitis patients with latex allergy.

    Science.gov (United States)

    Kimata, Hajime

    2003-01-01

    In atopic dermatitis patients with latex allergy, listening to Mozart reduced skin wheal responses induced by latex, but not by histamine, whereas listening to Beethoven failed to produce similar results. Listening to Mozart also decreased in vitro total IgE and latex-specific IgE production with concomitant skewing of the cytokine pattern toward the Th1 type, that is, an increase in Th1 cytokine production and decrease in Th2 cytokine production by peripheral blood mononuclear cells, whereas listening to Beethoven failed to do so. These results suggest that therapy using specific types of music may be an effective treatment of allergic diseases.

  9. A Novel Tricyclic Ligand-Containing Nonpeptidic HIV-1 Protease Inhibitor, GRL-0739, Effectively Inhibits the Replication of Multidrug-Resistant HIV-1 Variants and Has a Desirable Central Nervous System Penetration Property In Vitro

    Science.gov (United States)

    Amano, Masayuki; Tojo, Yasushi; Salcedo-Gómez, Pedro Miguel; Parham, Garth L.; Nyalapatla, Prasanth R.; Das, Debananda; Ghosh, Arun K.

    2015-01-01

    We report here that GRL-0739, a novel nonpeptidic HIV-1 protease inhibitor containing a tricycle (cyclohexyl-bis-tetrahydrofuranylurethane [THF]) and a sulfonamide isostere, is highly active against laboratory HIV-1 strains and primary clinical isolates (50% effective concentration [EC50], 0.0019 to 0.0036 μM), with minimal cytotoxicity (50% cytotoxic concentration [CC50], 21.0 μM). GRL-0739 blocked the infectivity and replication of HIV-1NL4-3 variants selected by concentrations of up to 5 μM ritonavir or atazanavir (EC50, 0.035 to 0.058 μM). GRL-0739 was also highly active against multidrug-resistant clinical HIV-1 variants isolated from patients who no longer responded to existing antiviral regimens after long-term antiretroviral therapy, as well as against the HIV-2ROD variant. The development of resistance against GRL-0739 was substantially delayed compared to that of amprenavir (APV). The effects of the nonspecific binding of human serum proteins on the anti-HIV-1 activity of GRL-0739 were insignificant. In addition, GRL-0739 showed a desirable central nervous system (CNS) penetration property, as assessed using a novel in vitro blood-brain barrier model. Molecular modeling demonstrated that the tricyclic ring and methoxybenzene of GRL-0739 have a larger surface and make greater van der Waals contacts with protease than in the case of darunavir. The present data demonstrate that GRL-0739 has desirable features as a compound with good CNS-penetrating capability for treating patients infected with wild-type and/or multidrug-resistant HIV-1 variants and that the newly generated cyclohexyl-bis-THF moiety with methoxybenzene confers highly desirable anti-HIV-1 potency in the design of novel protease inhibitors with greater CNS penetration profiles. PMID:25691652

  10. The organotypic multicellular spheroid is a relevant three-dimensional model to study adenovirus replication and penetration in human tumors in vitro

    NARCIS (Netherlands)

    Grill, Jacques; Lamfers, Martine L. M.; van Beusechem, Victor W.; Dirven, Clemens M.; Pherai, D. Shareen; Kater, Mathijs; van der Valk, Paul; Vogels, Ronald; Vandertop, W. Peter; Pinedo, Herbert M.; Curiel, David T.; Gerritsen, Winald R.

    2002-01-01

    The use of adenoviruses for gene transfer and as oncolytic agents is currently receiving widespread attention. As specific constraints to adenovirus distribution and spread cannot be studied in cell cultures, there is a need for an in vitro three-dimensional (3D) model mimicking the in vivo biology

  11. In Vitro susceptibility of Gram-positive cocci isolated from skin and respiratory tract to azithromycin and twelve other antimicrobial agents

    Directory of Open Access Journals (Sweden)

    Caio M. F. Mendes

    Full Text Available This study was conducted to evaluate the activity of azithromycin in comparison to 12 other antibacterial agents against recent isolates obtained consecutively from patients with respiratory tract or skin infections, from January to July, 2000. A total of 717 Gram-positive cocci were analyzed in this study and the following species were studied: Staphylococcus aureus (n=576, beta-hemolytic streptococci ( n=115, and Streptococcus pneumoniae (n=26. Susceptibility testing was carried out by the disk diffusion method and interpreted according to NCCLS breakpoints. The activity of azithromycin was compared to erythromycin, clindamycin, chloramphenicol, ciprofloxacin, ofloxacin, oxacillin, penicillin, ceftriaxone, tetracycline, trimethoprim/sulfamethoxazole, teicoplanin, and vancomycin. Of the 26 S. pneumoniae isolates recovered from the respiratory tract, 5 (19.2% were intermediate resistant to penicillin. All of these strains were susceptible to chloramphenicol, ofloxacin, and vancomycin, and 24 (92% were also susceptible to azithromycin, clindamycin, and erythromycin. Among the 67 beta-hemolytic streptococci strains isolated from the respiratory tract, 66 (99% were susceptible to azithromycin, erythromycin, clindamycin, and ofloxacin. All 48 beta-hemolytic streptococci strains isolated from skin were susceptible to azithromycin and clindamycin, 47 (98% were susceptible to erythromycin, and 46 (96% were susceptible to ofloxacin. Of the 576 strains of S. aureus, 253 (43.9% were isolated from the respiratory tract and 323 (56.1% from skin. Among S. aureus isolates from the respiratory tract and skin, 46 (18% and 78 (24%, respectively were resistant to oxacillin. Isolates from the respiratory tract and skin showed the same percentage of resistance (36% to azithromycin. These in vitro results suggest that azithromycin can be a therapeutic option for treatment of infections caused by these bacteria since the newer macrolides have several distinct

  12. Permeation of platinum and rhodium nanoparticles through intact and damaged human skin

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    Mauro, Marcella [University of Trieste, Clinical Unit of Occupational Medicine, Department of Medical Sciences (Italy); Crosera, Matteo; Bianco, Carlotta; Adami, Gianpiero; Montini, Tiziano; Fornasiero, Paolo [University of Trieste, Department of Chemical and Pharmaceutical Sciences (Italy); Jaganjac, Morana [Rudjer Boskovic Institute, Laboratory for Oxidative Stress, Department of Molecular Medicine (Croatia); Bovenzi, Massimo; Filon, Francesca Larese, E-mail: larese@units.it [University of Trieste, Clinical Unit of Occupational Medicine, Department of Medical Sciences (Italy)

    2015-06-15

    The aim of the study was to evaluate percutaneous penetration of platinum and rhodium nanoparticles (PtNPs: 5.8 ± 0.9 nm, RhNPs: 5.3 ± 1.9 nm) through human skin. Salts compounds of these metals are sensitizers and some also carcinogenic agents. In vitro permeation experiments were performed using Franz diffusion cells with intact and damaged skin. PtNPs and RhNPs, stabilized with polyvinylpyrrolidone, were synthesized by reduction of Na{sub 2}PtC{sub l6} and RhCl{sub 3}·3H{sub 2}O respectively. Suspensions with a concentration of 2.0 g/L of PtNPs and RhNPs were dispersed separately in synthetic sweat at pH 4.5 and applied as donor phases to the outer surface of the skin for 24 h. Measurements of the content of the metals in the receiving solution and in the skin were performed subsequently. Rhodium skin permeation was demonstrated through damaged skin, with a permeation flux of 0.04 ± 0.04 μg cm{sup −2} h{sup −1} and a lag time of 7.9 ± 1.1 h, while no traces of platinum were found in receiving solutions. Platinum and rhodium skin-analysis showed significantly higher concentrations of the metals in damaged skin. Rh and Pt applied as NPs can penetrate the skin barrier and Rh can be found in receiving solutions. These experiments pointed out the need for skin contamination prevention, since even a minor injury to the skin barrier can significantly increase penetration.

  13. Decolonisation of MRSA, S. aureus and E. coli by cold-atmospheric plasma using a porcine skin model in vitro.

    Directory of Open Access Journals (Sweden)

    Tim Maisch

    Full Text Available In the last twenty years new antibacterial agents approved by the U.S. FDA decreased whereas in parallel the resistance situation of multi-resistant bacteria increased. Thus, community and nosocomial acquired infections of resistant bacteria led to a decrease in the efficacy of standard therapy, prolonging treatment time and increasing healthcare costs. Therefore, the aim of this work was to demonstrate the applicability of cold atmospheric plasma for decolonisation of Gram-positive (Methicillin-resistant Staphylococcus aureus (MRSA, Methicillin-sensitive Staphylococcus aureus and Gram-negative bacteria (E. coli using an ex vivo pig skin model. Freshly excised skin samples were taken from six month old female pigs (breed: Pietrain. After application of pure bacteria on the surface of the explants these were treated with cold atmospheric plasma for up to 15 min. Two different plasma devices were evaluated. A decolonisation efficacy of 3 log(10 steps was achieved already after 6 min of plasma treatment. Longer plasma treatment times achieved a killing rate of 5 log(10 steps independently from the applied bacteria strains. Histological evaluations of untreated and treated skin areas upon cold atmospheric plasma treatment within 24 h showed no morphological changes as well as no significant degree of necrosis or apoptosis determined by the TUNEL-assay indicating that the porcine skin is still vital. This study demonstrates for the first time that cold atmospheric plasma is able to very efficiently kill bacteria applied to an intact skin surface using an ex vivo porcine skin model. The results emphasize the potential of cold atmospheric plasma as a new possible treatment option for decolonisation of human skin from bacteria in patients in the future without harming the surrounding tissue.

  14. Retinoic acid promotes the proliferation of primordial germ cell-like cells differentiated from mouse skin-derived stem cells in vitro.

    Science.gov (United States)

    Tan, Hui; Wang, Jun-Jie; Cheng, Shun-Feng; Ge, Wei; Sun, Yuan-Chao; Sun, Xiao-Feng; Sun, Rui; Li, Lan; Li, Bo; Shen, Wei

    2016-02-01

    Skin-derived stem cells (SDSCs) have the potential to differentiate into gametes and are a potential resource for research and clinical applications. Sufficient amount of primordial germ cells (PGCs) is an important requirement for successful differentiation of SDSCs into gametes in vitro. Retinoic acid (RA), a vitamin A-derived small lipophilic molecule, promotes the growth of PGCs in vivo; however, the role of RA on the proliferation of PGC-like cells (PGCLCs) derived from SDSCs remains unknown. In this study, SDSCs were induced to differentiate into the embryoid body and cocultured with mouse fibroblasts to form PGCLCs. The proliferation of PGCLCs with the presence of various concentrations of RA was investigated in vitro. Immunofluorescence labeling showed that the 5-Bromo-2-deoxyUridine-positive ratio of PGCLCs was increased after the cells were treated with 5-μM RA, and flow cytometry results showed that the number of cells in the S phase was increased significantly. The messenger RNA expression levels of cell cycle-related genes, CCND1 and CDK2, were also increased. Furthermore, RA effectively promoted the external proliferation of endogenous PGCs when 11.5-days postcoitum fetal mouse genital ridges were cultured in vitro. In conclusion, 5-μM RA promoted the proliferation of SDSCs-derived PGCLCs and endogenous PGCs. Our study will provide a valuable model system for studying the differentiation of stem cells into gametes in vitro. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Corneocyte quantification by NIR densitometry and UV/Vis spectroscopy for human and porcine skin and the role of skin cleaning procedures.

    Science.gov (United States)

    Schwarz, J C; Klang, V; Hoppel, M; Wolzt, M; Valenta, C

    2012-01-01

    Optical methods of corneocyte quantification during tape stripping experiments on the skin are useful tools for the rapid evaluation of the skin penetration potential of dermally applied substances. However, a comparative investigation of the different methods proposed for this task, namely NIR densitometry and UV/Vis spectroscopy, is still missing. Thus, the aim of the present work was to employ these two techniques in comparative tape stripping experiments both in vivo on human forearm skin and in vitro on porcine ear skin. Standard tape stripping experiments were performed in the absence and presence of a marketed formulation containing flufenamic acid as a model drug. In the context of these methodological investigations, different methods of skin cleaning prior to the tape stripping procedure were evaluated to identify the most appropriate working protocol among the approaches proposed in the respective literature. The results showed that the investigated methods of NIR densitometry and UV/Vis spectroscopy deliver highly comparable results. Both optical methods are suitable to determine the skin penetration profiles of active substances during in vivo and in vitro tape stripping, especially if a simple working protocol without any cleaning procedures is maintained. Copyright © 2012 S. Karger AG, Basel.

  16. A comparative study of baby immature and adult shoots of Aloe vera on UVB-induced skin photoaging in vitro.

    Science.gov (United States)

    Hwang, Eunson; Kim, Su Hyeon; Lee, Sarah; Lee, Choong Hwan; Do, Seon-Gil; Kim, Jinwan; Kim, Sun Yeou

    2013-12-01

    Ultraviolet (UV) irradiation induces photo-damage of the skin, which in turn causes depletion of the dermal extracellular matrix and chronic alterations in skin structure. Skin wrinkle formations are associated with collagen synthesis and matrix metalloproteinase (MMP) expression. The production of type I procollagen is regulated by transforming growth factor-β1 (TGF-β1) expression; the activation of MMP is also correlated with an increase of interleukin-6 (IL-6). Aloe barbadensis M. (Aloe vera) is widely used in cosmetic and pharmaceutical products. In this study, we examined whether baby aloe shoot extract (BAE, immature aloe extract), which is from the one-month-old shoots of Aloe vera, and adult aloe shoot extract (AE), which is from the four-month-old shoots of Aloe vera, have a protective effect on UVB-induced skin photoaging in normal human dermal fibroblasts (NHDFs). The effects of BAE and AE on UVB-induced photoaging were tested by measuring the levels of reactive oxygen species, MMP-1, MMP-3, IL-6, type I procollagen, and TGF-β1 after UVB irradiation. We found that NHDF cells treated with BAE after UVB-irradiation suppressed MMP-1, MMP-3, and IL-6 levels compared to the AE-treated cells. Furthermore, BAE treatment elevated type I procollagen and TGF-β1 levels. Our results suggest that BAE may potentially protect the skin from UVB-induced damage more than AE. Copyright © 2013 John Wiley & Sons, Ltd.

  17. Effects of plant sterols derived from Aloe vera gel on human dermal fibroblasts in vitro and on skin condition in Japanese women

    Directory of Open Access Journals (Sweden)

    Tanaka M

    2015-02-01

    Full Text Available Miyuki Tanaka,1 Eriko Misawa,1 Koji Yamauchi,1 Fumiaki Abe,1 Chiaki Ishizaki2 1Functional Food Research Department, Food Science and Technology Institute, Morinaga Milk Industry Co, Ltd, Zama, Kanagawa, 2Ebisu Skin Research Center, Inforward, Inc., Tokyo, Japan Background: Aloe is known for its topical use for treating wounds and burns. Many previous studies reported the healing effects of Aloe vera. However, there are few clinical studies on the effect of orally administered A. vera gel on the skin. Aloe sterols are a type of plant sterols that have the capability to regulate the metabolism of glucose and lipids. In a recent study, we confirmed that ingested Aloe sterols reached the peripheral tissues through the bloodstream. However, their influence on dermal fibroblasts has not been investigated. Methods: First, we investigated the capability of Aloe sterols (cycloartenol and lophenol to stimulate human dermal fibroblasts in vitro. Then, we investigated the effect of intake of Aloe vera gel powder (AVGP containing 40 µg Aloe sterols on the skin conditions in Japanese women with dry skin in a randomized, double-blind, placebo-controlled trial. Results: After cocultivation with Aloe sterols, the production of collagen and hyaluronic acid increased by approximately two-fold and 1.5-fold, and gene expression levels of these enzymes responsible for their synthesis were also observed in human dermal fibroblasts. An increase in arm skin hydration was observed at 8 weeks in the AVGP group, whereas a slight decrease in arm skin hydration was noted in the placebo group. However, there was no statistical difference between AVGP and placebo groups in skin moisture. In subgroup analysis, the change in the mean wrinkle depth was significantly lower in the AVGP group than in the control group. In addition, percent body fat after 8 weeks was significantly lower in the AVGP group. No AVGP intake-dependent harmful phenomenon was observed during the intake

  18. In vitro assessment of eye irritancy using the Reconstructed Human Corneal Epithelial SkinEthic HCE model: application to 435 substances from consumer products industry.

    Science.gov (United States)

    Cotovio, José; Grandidier, Marie-Hélène; Lelièvre, Damien; Bremond, Christelle; Amsellem, Carolle; Maloug, Saber; Ovigne, Jean-Marc; Loisel-Joubert, Sophie; Lee, Aline Van Der; Minondo, Anne-Marie; Capallere, Christophe; Bertino, Béatrice; Alépée, Nathalie; Tinois-Tessonneaud, Estelle; de Fraissinette, Anne De Brugerolle; Meunier, Jean-Roch; Leclaire, Jacques

    2010-03-01

    The 7th amendment of the EU Cosmetics Directive led to the ban of eye irritation testing for cosmetic ingredients in animals, effective from March 11th 2009. Over the last 20years, many efforts have been made to find reliable and relevant alternative methods. The SkinEthic HCE model was used to evaluate the in vitro eye irritancy potential of substances from a cosmetic industry portfolio. An optimized protocol based on a specific 1-h treatment and a 16-h post-treatment incubation period was first assessed on a set of 102 substances. The prediction model (PM) based on a 50% viability cut-off, allowed to draw up two classes (Irritants and Non-Irritants), with good associated sensitivity (86.2%) and specificity (83.5%). To check the robustness of the method, the evaluated set was expanded up to 435 substances. Final performances maintained a high level and were characterized by an overall accuracy value > 82% when using EU or GHS classification rules. Results showed that the SkinEthic HCE test method is a promising in vitro tool for the prediction of eye irritancy. Optimization datasets were shared with the COLIPA Eye Irritation Project Team and ECVAM experts, and reviewed as part of an ongoing progression to enter an ECVAM prospective validation study for eye irritation. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

  19. The skin-depigmenting potential of Paeonia lactiflora root extract and paeoniflorin: in vitro evaluation using reconstructed pigmented human epidermis.

    Science.gov (United States)

    Qiu, J; Chen, M; Liu, J; Huang, X; Chen, J; Zhou, L; Ma, J; Sextius, P; Pena, A-M; Cai, Z; Jeulin, S

    2016-10-01

    The roots of the herb Paeonia lactiflora ('White Peony') are used in association with other herbs in traditional clinical cosmetic practice in China as oral treatment for skin pigmentary disorders, such as brown or dark pigmentary spots. However, the skin-depigmenting potential of Paeonia lactiflora root extract and its main ingredient paeoniflorin has been scarcely investigated by topical application. The purpose of this study was to evaluate the efficacy of Paeonia lactiflora root extract and paeoniflorin as skin whitening agent in cosmetic application. Paeonia lactiflora root extract (containing 53.25% of paeoniflorin) and paeoniflorin (97% purity) were applied topically on reconstructed pigmented human epidermis model, a three-dimensional (3D) human skin equivalent, showing morphological and functional characteristics similar to those of in vivo human skin. Two specific methods were used for quantifying melanin inside the reconstructed pigmented epidermis: Fontana-Masson staining (2D quantification) and multiphoton microscopy (3D quantification). Compared to vehicle (dimethyl sulfoxide DMSO), a significant decrease in 2D and 3D melanin content was observed after topical application on reconstructed pigmented epidermis of Paeonia lactiflora extract at 300 μg mL(-1) (-28% and -27%, respectively) and paeoniflorin at 120 μg mL(-1) /250 μM (-30% and -23%, respectively), which is in the same order of magnitude as the positive reference 4-n-butylresorcinol at 83 μg mL(-1) /500 μM (-26% and -40%, respectively). These results demonstrate, for the first time, the depigmenting potential of paeoniflorin and thus the potential interest of using Paeonia lactiflora root extracts containing paeoniflorin in cosmetic or dermatological applications for reducing the severity of some hyperpigmented skin disorders. © 2016 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  20. Catch-up validation study of an in vitro skin irritation test method based on an open source reconstructed epidermis (phase I).

    Science.gov (United States)

    Mewes, K R; Fischer, A; Zöller, N N; Laubach, V; Bernd, A; Jacobs, A; van Rompay, A; Liebsch, M; Pirow, R; Petersohn, D

    2016-10-01

    We have developed a new in vitro skin irritation test based on an open source reconstructed epidermis (OS-REp) with openly accessible protocols for tissue production and test performance. Due to structural, mechanistic and procedural similarity, a blinded catch-up validation study for skin irritation according to OECD Performance Standards (PS) was conducted in three laboratories to promote regulatory acceptance, with OS-REp models produced at a single production site only. While overall sensitivity and predictive capacity met the PS requirements, overall specificity was only 57%. A thorough analysis of the test results led to the assumption that some of the false-positive classifications could have been evoked by volatile skin-irritating chemicals tested in the same culture plate as the non-irritants falsely predicted as irritants. With GC/MS and biological approaches the cross-contamination effect was confirmed and the experimental set-up adapted accordingly. Retesting of the affected chemicals with the improved experimental set-up and otherwise identical protocol resulted in correct classifications as non-irritants. Taking these re-test results into account, 93% overall sensitivity, 70% specificity and 82% accuracy was achieved, which is in accordance with the OECD PS. A sufficient reliability of the method was indicated by a within-laboratory-reproducibility of 85-95% and a between-laboratory-reproducibility of 90%. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. Catch-up validation study of an in vitro skin irritation test method based on an open source reconstructed epidermis (phase II).

    Science.gov (United States)

    Groeber, F; Schober, L; Schmid, F F; Traube, A; Kolbus-Hernandez, S; Daton, K; Hoffmann, S; Petersohn, D; Schäfer-Korting, M; Walles, H; Mewes, K R

    2016-10-01

    To replace the Draize skin irritation assay (OECD guideline 404) several test methods based on reconstructed human epidermis (RHE) have been developed and were adopted in the OECD test guideline 439. However, all validated test methods in the guideline are linked to RHE provided by only three companies. Thus, the availability of these test models is dependent on the commercial interest of the producer. To overcome this limitation and thus to increase the accessibility of in vitro skin irritation testing, an open source reconstructed epidermis (OS-REp) was introduced. To demonstrate the capacity of the OS-REp in regulatory risk assessment, a catch-up validation study was performed. The participating laboratories used in-house generated OS-REp to assess the set of 20 reference substances according to the performance standards amending the OECD test guideline 439. Testing was performed under blinded conditions. The within-laboratory reproducibility of 87% and the inter-laboratory reproducibility of 85% prove a high reliability of irritancy testing using the OS-REp protocol. In addition, the prediction capacity was with an accuracy of 80% comparable to previous published RHE based test protocols. Taken together the results indicate that the OS-REp test method can be used as a standalone alternative skin irritation test replacing the OECD test guideline 404. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. Analysis of the penetration of a caffeine containing shampoo into the hair follicles by in vivo laser scanning microscopy

    Science.gov (United States)

    Lademann, J.; Richter, H.; Schanzer, S.; Klenk, A.; Sterry, W.; Patzelt, A.

    2010-02-01

    In previous in vitro investigations, it was demonstrated that caffeine is able to stimulate the hair growth. Therefore, a penetration of caffeine into the hair follicle is necessary. In the present study, in vivo laser scanning microscopy (LSM) was used to investigate the penetration and storage of a caffeine containing shampoo into the hair follicles. It was shown that a 2-min contact time of the shampoo with the skin was enough to accumulate significant parts of the shampoo in the hair follicles. A penetration of the shampoo up to a depth of approx. 200 μm could be detected, which represents the detection limit of the LSM. At this depth, the close network of the blood capillaries surrounding the hair follicles commences. Even after 24 h, the substance was still detectable in the hair follicles. This demonstrates the long-term reservoir function of the hair follicles for topically applied substances such as caffeine.

  3. A comparison of the resin tag penetration of the total etch and the self-etch dentin bonding systems in the primary teeth: An in vitro study

    Directory of Open Access Journals (Sweden)

    Sajjad Mithiborwala

    2012-01-01

    Full Text Available Background and Objective : Restoration of carious lesions with a strong permanent bond would be a highly desirable requisite. Ultra morphological characterization shows that observing and understanding the interfacial phenomenon and its quality would be of great importance in the selection of a dental adhesive for its use in pediatric restorative dentistry. Study design : Human primary molars, indicated for extraction, for reasons like caries, normal exfoliation, pathological root resorption, over-retained and serial extraction, were collected. Teeth were then equally distributed into 2 subgroups each namely B1 - Prime and Bond NT & B2 - Xeno III. Results : The resin tags seen in the samples of group B2 were both qualitatively and quantitatively advanced as compared to group B1. This reveals that the quality of the penetration of the resin was better in group B2. Conclusion : Reduction in the technique sensitivity of any bonding system would always be a preferred factor in pediatric restorative dentistry. Thus the inclination towards the selection of adhesive system may lean towards the self-etching bonding system at this juncture.

  4. Effect of olive oil on transdermal penetration of flurbiprofen from topical gel as enhancer.

    Science.gov (United States)

    Hussain, Abid; Khan, Gul Majid; Jan, Syed Umer; Shah, Shefaatullah; Shah, Kifayatullah; Akhlaq, Muhammad; Rahim, Nouman; Nawaz, Asif; Wahab, Abdul

    2012-04-01

    The present study was conducted to formulate and evaluate flurbiprofen transdermal gel. A standard calibration curve was constructed to obtain a regression line equation to be used for finding out the concentration of drug i