Acne is a multifactorial skin disease affecting about 80 % of people aged 11 to 30. Several systemic and topical treatments are used to treat existing lesions, prevent scarring and suppress the development of new lesions. Topical therapy is often used as first line treatment for acne, due to the location of the target organ, the pilosebaceous unit, in the skin. Retinoids are widely used as oral or topical treatment for this disease, with tretinoin and adapalene being two of the most used topical retinoids. The transdermal route offers several challenges to drug delivery, e.g. the excellent resistance of the stratum corneum to diffusion, as well as variable skin properties such as site, age, race and disease. Some additional difficulties are associated with the dermatological delivery of tretinoin and adapalene, which include suboptimal water solubility of the retinoids, isomerisation of tretinoin in the skin, mild to severe skin irritation, as well as oxidation and photo–isomerisation of tretinoin, even before crossing the stratum corneum. Researchers constantly strive to improve dermatological retinoid formulations in order to combat low dermal flux, skin irritation and instability. The release kinetics of tretinoin varies greatly according to the way in which it is incorporated into the formulation and according to the type of formulation used. Little research has been conducted regarding improved formulations for adapalene. Pheroid technology is a patented delivery system employed in this study in order to improve the dermal delivery of retinoids. Tretinoin and adapalene were separately incorporated into castor oil, vitamin F and Pheroid creams. The creams were evaluated in terms of their in vitro retinoid release, in vitro transdermal diffusion and stability. Castor oil and Pheroid creams were superior in terms of release and dermal delivery of adapalene. Tretinoin was best released and delivered to the dermis by castor oil cream. The castor oil creams were the most stable formulations, whereas the Pheroid creams were the most unstable. In terms of release, dermal diffusion and stability, castor oil cream proved to be the most suitable cream for both tretinoin and adapalene. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2011.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Technology Candidate: Pavel Hladký Consultant: PharmDr. Barbora Švecová, Ph.D. Title of Thesis: Formulation and (trans)dermal application of imiquimod Imiquimod (IMQ) is an active pharmaceutical substance which belongs to the group of heterocyclic imidazoquinolines. The mechanism of its effect is an induction of cellular immune response after topical administration, that is used for a treatment of tumors or viral diseases of the skin. In the Czech Republic it is available like a cream called Aldara® , the content of IMQ is 5 %. Although Aldara® is an effective medicine, many problems are associated with its use, especially high price, undesirable effects, disposable use, environmental pollution, etc. The aim of this work was to prepare new liposomes for topical administration containing lower ammount of IMQ (0.5 %) and evaluation of penetration of IMQ into human skin in vitro. To improve the entrance of the drug into the skin transdermal penetration enhancers were used. Permeation experiments were performed in Franz diffusion cells on human skin under conditions as close as possible to the physiological environment of the organism. Subsequently, the individual layers of the skin (stratum corneum, the epidermis,...

Made available in DSpace on 2014-06-11T19:25:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-21Bitstream added on 2014-06-13T20:33:03Z : No. of bitstreams: 1 martini_pc_me_arafcf.pdf: 595100 bytes, checksum: bcb255af52349311710acfecbb49c0e8 (MD5) / Universidade Estadual Paulista (UNESP) / O aumento da expectativa de vida tem provocado grande demanda por produtos que auxiliem na prevenção do envelhecimento da pele. O ácido hialurônico (AH) está sendo muito utilizado em formulações antienvelhecimento por se acreditar que ele possa atenuar os efeitos que o tempo produz na pele. Porém, para o desenvolvimento de um cosmético, é necessário avaliar o risco potencial dos ingredientes que compõem a formulação, que devem estar em concentração que apresente margem de segurança adequada, sendo importante a realização de teste de absorção, estudo do potencial de risco irritativo e testes de mutagenicidade. O objetivo deste estudo foi desenvolver um gel de ácido hialurônico com alta massa molecular e incorporar ácido hialurônico de massas moleculares menores, realizar o controle microbiológico dos géis, avaliar a toxicidade dérmica aguda e a citotoxicidade, determinar o comportamento reológico das formulações, analisar seu perfil de liberação, permeação e retenção cutânea in vitro. Foi preparado como gel base, a mistura de água e AH de alta massa molecular e, posteriormente, neste gel formado foram incorporados os AHs de menores massas moleculares. O controle de qualidade microbiológico foi realizado de acordo com a Farmacopeia Brasileira (2010), a avaliação da toxicidade dérmica aguda foi realizada utilizando-se 25 ratos wistar, que foram divididos em 5 grupos com 5 animais em que cada grupo recebeu a aplicação de um dos géis para posterior analise dos resultados. Para a avaliação da citotoxicidade utilizou-se o método colorimétrico 3-(4,5-dimetiltiazol-2-il)2,5-difenil brometo de tetrazoilium (MTT) descrito por MOSMANN, utilizando-se linhagens celulares de HepG2 (Human Epidermoide Cancer Cells) e HaCaT. As amostras tiveram seu comportamento reológico avaliado em reômetro HAAKE. O estudo de liberação, permeação e retenção cutânea... / Increased life expectancy has caused a great demand for products that help in the prevention of skin aging. Hyaluronic acid is being used in anti-aging formulations because it is expected that it can decrease the effects of time in the skin. However, for the development of a cosmetic it is necessary to evaluate the potential risk of the ingredients in the formulation that should be with adequate concentration to provide a margin of safety, being important to carry out absorption test, studies of the potential risk of irritating the skin and mutagenicity tests. The aim of this study was to develop a gel of hyaluronic acid with high molecular weight and incorporate hyaluronic acid with small molecular weights in it, analyze the microbiological control of the gels, evaluate the acute dermal toxicity and cytotoxicity, determine rheological characteristics, analyze the release profile, skin permeation and retention in vitro. It was prepared like gel base, the mixture of water and AH of high molecular weight, and after was incorporate AH of smaller molecular weight. The quality control microbiological was accomplished in agreement with Brazilian Pharmacopoeia (2010), the evaluate the acute dermal toxicity was made with 25 rat wistar, that were divided in 5 groups with 5 animals in each group received the application from one of the gels for subsequent analyzes of the results. For the evaluation of the cytotoxicity was used the method colorimeter 3-(4,5-dimetiltiazol-2-il)2,5-difenil tetrazoilium bromide (MTT) described by MOSMANN (1983), being used cellular lineages of HepG2 (Human Epidermoide Câncer Cells) and HaCaT. The samples had its behavior rheological characteristics. The analyze the release profile, skin permeation and retention were accomplished using cells of Franz modified. The microbiological was in agreement... (Complete abstract click electronic access below)

In our modern society we are exposed to many natural and synthetic chemicals. The assessment of chemicals with regard to human safety is difficult but nevertheless of high importance. Beside clinical studies, which are restricted to potential pharmaceuticals only, most toxicity data relevant for regulatory decision-making are based on in vivo data. Due to the ban on animal testing of cosmetic ingredients in the European Union, alternative approaches, such as in vitro and in silico tests, have become more prevalent. In this thesis existing non-testing approaches (i.e. studies without additional experiments) have been extended, e.g. QSAR models, and new non-testing approaches, e.g. in vitro data supported structural alert systems, have been created. The main aspect of the thesis depends on the determination of data quality, improving modelling performance and supporting Adverse Outcome Pathways (AOPs) with definitions of structural alerts and physico-chemical properties. Furthermore, there was a clear focus on the transparency of models, i.e. approaches using algorithmic feature selection, machine learning etc. have been avoided. Furthermore structural alert systems have been written in an understandable and transparent manner. Beside the methodological aspects of this work, cosmetically relevant examples of models have been chosen, e.g. skin penetration and hepatic steatosis. Interpretations of models, as well as the possibility of adjustments and extensions, have been discussed thoroughly. As models usually do not depict reality flawlessly, consensus approaches of various non-testing approaches and in vitro tests should be used to support decision-making in the regulatory context. For example within read-across, it is feasible to use supporting information from QSAR models, docking, in vitro tests etc. By applying a variety of models, results should lead to conclusions being more usable/acceptable within toxicology. Within this thesis (and associated publications) novel methodologies on how to assess and employ statistical data quality and how to screen for potential liver toxicants have been described. Furthermore computational tools, such as models for skin permeability and dermal absorption, have been created.

O surgimento das matrizes de regeneração dérmica nas duas últimas décadas permitiu um grande avanço no tratamento tanto das queimaduras agudas como das sequelas. No entanto, ainda há carência de informações sobre a relação entre os resultados clínicos e o que ocorre no tecido com cada tipo de matriz. O objetivo deste estudo foi avaliar prospectivamente os aspectos clínicos quanto à qualidade de pele, escala de Vancouver e POSAS, função e retração da área tratada e os aspectos histológicos na microscopia de luz e eletrônica, com o uso de duas matrizes de regeneração dérmica, ambas de colágeno bovino, uma de duas camadas, recoberta com lâmina de silicone e outra sem. Vinte e quatro pacientes, sorteados 12 em cada grupo, tiveram suas retrações cicatriciais secundárias à queimaduras tratadas em duas cirurgias, a primeira de liberação da retração e colocação da matriz e a segunda, colocação do auto enxerto de pele; em ambas as cirurgias foi utilizado o curativo de pressão negativa. As avaliações da escala de Vancouver e medidas da retração da área foram realizadas no pré-operatório, 1, 3, 6 e 12 meses e a escala de POSAS e avaliação funcional no pré-operatório e aos 12 meses. As biópsias foram colhidas no pré-operatório, no dia da colocação do enxerto de pele, 12 dias, 2, 6 e 12 meses após o enxerto. A avaliação clínica mostrou retração de todas as áreas tratadas, melhora da qualidade da pele e funcional em todos os pacientes. A matriz com silicone, mostrou superioridade dos resultados quanto a qualidade da pele, função e menor retração da área tratada. A análise histológica mostrou o crescimento de tecido conjuntivo denso idêntico ao tecido cicatricial original, sem diferenças entre as matrizes e que não se assemelha à derme normal. Também não foi observada diferença no diâmetro das fibrilas de colágeno do tecido neoformado, a pele normal e a cicatriz / The advent of dermal regenerate templates has fostered major advances in the treatment of acute burns and their sequelae, in the last two decades. Both data on morphological aspects of the newly-formed tissue, and clinical trials comparing different templates, are still lacking. The goal of this study was to prospectively analyze the outcome of patients treated with two of the existing templates, followed by thin skin autograft. They are both made of bovine collagen, one includes a superficial silicone layer. Surgery was performed on patients with impaired mobility resulting from burn sequelae (n = 12 per template). Negative pressure therapy was applied post-surgically; patients were monitored for 12 months. Data on scar skin quality (Vancouver and POSAS evaluation scales), rate of joint mobility recovery, and graft contraction were recorded; as well as morphological analyses at light microscopical and ultrastructural levels. Improvement in mobility and skin quality were demonstrated along with graft contraction, in all patients. The silicone-coupled template showed the best performance in all aspects. There was sub epidermal growth of dense connective tissue, indistinguishable from the original scars in both templates. The formation of tissue resembling normal dermis was not detected in any of the cases. Likewise, the ultrastructural analysis showed the same architecture of the connective tissue among the template scars and the original scar. No difference was detected when the collagen fibril diameters of the normal skin and of the scars (original and of the two templates) were compared

Dysregulated wound healing is commonly associated with excessive fibrosis. Connective tissue growth factor (CTGF/CCN2) is characteristically overexpressed in fibrotic diseases and stimulated by transforming growth factor-β (TGF-β) in dermal fibroblasts. Reepithelialisation and epidermal wound coverage counteract excessive scar formation. We have previously shown that interleukin-1α (IL-1α) derived from keratinocytes conteracts TGF-β-stimulated CTGF-expression. The aim of this thesis was to further explore the effects of keratinocytes and IL-1α on gene and protein expression, as well as pathways, in TGF-β stimulated fibroblasts. Fibroblasts were studied in vitro by conventional two dimensional cell culture models and in a three dimensional keratinocyte-fibroblast organotypic skin culture model. The results showed that IL-1 suppresses basal and TGF-β-induced CTGF mRNA and protein, involving a possible TAK1 mechanism. Keratinocytes regulate the expression of fibroblast genes important for the turnover of the extracellular matrix. Most of the genes analysed (11/13) were regulated by TGF-β and counter regulated by keratinocytes. The overall results support a view that keratinocytes regulate fibroblasts to act catabolically (anti-fibrotic) on the extracellular matrix. Transcriptional microarray and gene set enrichment analysis showed that antagonizing effects of IL-1α on TGF-β were much more prominent than the synergistic effects. The most confident of these pathways was the interferon signaling, which were inhibited by TGF-β and activated by IL-1α. A proteomics study confirmed that IL-1α preferentially conteracts TGF-β effects. Six new fibroblast proteins involved in synthesis/ regulation were identified, being regulated by TGF-β and antagonized by IL-1α. Pathway analysis confirmed counter-regulation of interferon signaling by the two cytokines. These findings have implications for understanding the role of fibroblasts for inflammatory responses and development of fibrosis in the skin.

No description available.

No description available.

No description available.

Radiotherapy has major therapeutic benefits for cancer patients, but ionizing radiation causes damage of surrounding healthy tissues with poor wound healing a common side effect. Therefore, further oncoplastic, reconstructive surgery is challenging and often problematic. Current research models use normal human dermal fibroblasts irradiated in vitro to mimic radiation damage, but this is not comparable to ionising radiation and only measures acute changes. Since radiotherapy may induce epigenetic changes leading to alterations in dermal fibroblast phenotype, the first aim of this study was to compare fibroblasts cultured from irradiated skin with non-irradiated skin. As mesenchymal stem cells isolated from adipose tissue may offer beneficial effects in the regenerative capacity of irradiated tissue, the second part of this study was to compare those cultured from non-irradiated and irradiated breast tissue. Histological changes in the structural organisation of breast tissue in situ from donors exposed to radiotherapy was compared to untreated breast. Primary cultures of dermal fibroblasts from irradiated and non-irradiated breast skin were established and comparisons quantitated in proliferation (CyQuant), metabolism (Alamar Blue), migration (scratch-wound assay), collagen production (Sircol), levels of proteases and protease inhibitors (human protease/protease inhibitor array) and gene expression of COL1A1, COL3A1, MMP1, MMP2, TIMP1 and PPAR-γ mRNA (qPCR). Cells from the stromal vascular fraction (SVF) were cultured and characterised by immunocytochemistry and compared to human preadipocytes sourced commercially. The secretion of FGF, adiponectin and VEGF by the preadipocyte and the SVF mesenchymal cells was compared and the ability of their secretome to modulate dermal fibroblast proliferation, metabolism and migration was evaluated. Radiotherapy caused extensive disorganisation of the reticular dermis and flattening of the epidermal-dermal junction. Dermal fibroblasts cultured from irradiated skin had a pronounced spindle shaped morphology with longer thinner projections and took approximately twice as long to explant and grow. They had a lower proliferative and higher basal metabolic rate and did not respond to FGF-2. While they secreted similar amounts of total collagen they demonstrated distinct differences in proteolytic enzyme and protease inhibitor expression. This is the first report to culture cells from the SVF of irradiated breast tissue. The cells expressed the preadipocyte markers CD10, CD73 and CD105 and no CD45 (negative marker). SVF cells cultured displayed a typical ASC fibroblastoid morphology. Analysis of the secretome identified the presence of FGF, adiponectin and VEGF, while functional analysis demonstrated a stimulatory effect on normal dermal fibroblast migration, although irradiated dermal fibroblasts were unresponsive. Radiotherapy induces long term, detrimental changes in breast skin. This is the first quantitative characterisation of dermal fibroblasts and mesenchymal cells from the SVF, subjected to ionising radiation in situ. Changes in their phenotype that alter their function will impact on wound healing. Further characterisation of these cells may explain their dysfunctional behaviour, and lead to therapies to reverse or reduce this deleterious side-effect and significantly improve treatments facilitating wound healing following radiation injury. / Plastic Surgery and Burns Research unit