Um dos tipos de câncer mais frequentes no mundo é o carcinoma epidermoide de cabeça e pescoço. A dificuldade de resposta eficiente aos tratamentos está relacionada à complexidade molecular deste tumor. MicroRNAs regulam uma parte significativa dos genes humanos, dentre os quais genes responsáveis por controlar proliferação e apoptose, caracterizando seu papel na progressão tumoral e resistência a quimioterápicos. O entendimento sobre como moléculas com ação antineoplásica atuam sobre a regulação de microRNAs pode contribuir para a compreensão de resultados terapêuticos. Neste estudo avaliamos o efeito dos triterpenos quinonametídeos maitenina e 22-β-hidroximaitenina sobre a viabilidade celular e sobre a regulação de microRNAs associados com proliferação e apoptose. O efeito destas moléculas foi avaliado frente a linhagens tumorais e em queratinócitos orais livres de tumor. Os resultados indicam que os dois triterpenos avaliados reduzem eficazmente a viabilidade das células cancerígenas e a regulação de microRNAs faz parte dos mecanismos envolvidos neste efeito. / Head and neck squamous cell carcinoma is one of the most frequent cancer types in the world. The difficulty of effective response to treatment is related to the molecular complexity of this tumor. MicroRNAs, small non-coding RNA molecules, regulate a significant proportion of human genes, among which genes involved in cell growth, apoptosis and response to chemotherapy. The comprehension on how anticancer molecules interfere with microRNA expression should contribute to the understanding of treatment outcome. This study evaluated the effect of two quinonamethide triterpenoids, maytenin and 22-β-hidroxymaytenin, on cell viability and on the regulation of microRNAs associated with proliferation and apoptosis. The effect of these molecules was evaluated in tumor cells and oral keratinocytes. The results show that the two triterpenoids effectively reduced cell viability of cancer-derived cell lines and that microRNA regulation is one of the mechanisms involved in this effect.

No description available.

Autosomal dominant epidermolytic ichthyosis (EI) is a rare disease characterized by intra-epidermal blistering due to mutations in either of two keratin genes, KRT1 and KRT10, expressed by suprabasal keratinocytes. Autosomal recessive congenital ichthyosis (ARCI) is a non-blistering, hyperkeratotic disease caused by mutations in one of the following genes: ABCA12, ALOX12B, ALOXE3, TGM1, CYP4F22, NIPAL4 and SLC27A4, which are all essential for skin barrier homeostasis. ARCI and EI often respond well to treatment with retinoids, but the mechanism of action is unclear. The aim of this thesis was to increase the knowledge of pathogenic pathways in ichthyosis and to find new explanations to the effect of retinoids. In vitro studies of immortalized keratinocytes from EI patients showed an abnormal keratin aggregation after heat stress, that could be partially inhibited by pre-treatment with all-trans retinoic acid (ATRA) or retinoic acid receptor α-agonists. ATRA treatment also reduced the relative expression of mutated vs wildtype KRT10. The clearance of ATRA in human keratinocytes was found to be mediated by CYP26B1. In skin biopsies from ARCI patients, immunofluorescence analysis of 12R-LOX, eLOX-3, TGM1, ichthyin and FATP4 showed altered expression, not only of the mutated protein, but also of the other proteins. These observations are consistent with a feedback regulatory mechanism by which the loss of one protein results in an up-regulation of other proteins. Furthermore, 12R-LOX, eLOX-3 and TGM1 were intimately co-localized in stratum corneum, as were ichthyin and FATP4, suggesting that the proteins are linked to the same metabolic pathway. When treated with a CYP26 inhibitor known to raise the endogenous ATRA level of the skin, two patients with NIPAL4 mutations, initially exhibiting increased co-localization signals for 12R-LOX and eLOX-3, displayed normalized lipoxygenase expressions and showed clinical improvement. In conclusion, mechanisms are proposed by which pathogenic keratin aggregations in EI and epidermal protein deficiencies in ARCI patients may be mitigated by retinoids. Furthermore, the vivid crosstalk between proteins incriminated in ARCI suggests that these enzymes operate along a common metabolic pathway essential for producing barrier lipids in stratum corneum. Any abrogation of this production may cause barrier failure, hence resulting in a compensatory hyperkeratosis characteristic of congenital ichthyosis.

Chemische und physikalische Eigenschaften von Polymeren können verschiedene Zelltypen unterschiedlich, z. B. hinsichtlich Adhärenz oder Funktionalität, beeinflussen. Die Elastizität eines Polymers beeinflusst vor allem, welche Zugkräfte eine Zelle gegenüber ihrem Substrat entwickeln kann. Das Zellverhalten wird dann über intrazelluläre Rückkopplungsmechanismen reguliert. Die Oberflächenladung und/oder Hydrophilie eines Polymers beeinflusst zunächst die Adsorption von Ionen, Proteinen und anderen Molekülen. Vor allem über die Zusammensetzung, Dichte und Konformation der adsorbierten Komponenten werden anschließend die Wechselwirkungen mit den Zellen vermittelt. Des Weiteren können verschiedene Zelltypen unterschiedliche membranassoziierte Proteine, Zucker und Lipide aufweisen, so dass Polymereigenschaften zellspezifische Effekte bewirken können. Für biotechnologische Anwendungen und für den Einsatz in der regenerativen Medizin gewinnen Polymere, die spezifische Zellreaktionen regulieren können, immer weiter an Bedeutung. Die Isolierung und Kultur von primären Keratinozyten ist noch immer anspruchsvoll und die adäquate Heilung von Hautwunden stellt eine fortwährende medizinische Herausforderung dar. Ein Polymer, das eine bevorzugte Adhärenz von Keratinozyten bei gleichzeitig verminderter Anheftung dermaler Fibroblasten ermöglicht, würde erhebliche Vorteile für den Einsatz in der Keratinozyten-Zellkultur und als Wundauflage bieten. Um den potentiell spezifischen Einfluss bestimmter Polymereigenschaften auf primäre humane Keratinozyten und dermale Fibroblasten zu untersuchen, wurde in der vorliegenden Arbeit ein Zellkultursystem für die Mono- und Cokultur beider Zelltypen entwickelt. Das Testsystem wurde als Screening konzipiert, um den Einfluss unterschiedlicher Polymereigenschaften in mehreren Abstufungen auf die Zellen zu untersuchen. Folgende Parameter wurden untersucht: 1. Vitalität und Dichte adhärenter und nicht-adhärierter Zellen, 2. Schädigung der Zellmembran, 3. selektive Adhärenz von Keratinozyten in Cokultur durch die spezifische immunzytochemische Färbung von Keratin14 und Vimentin. Für die Polymere mit variabler Elastizität wurden zusätzlich die Ablagerung extrazellulärer Matrixkomponenten und die Sekretion löslicher Faktoren durch die Zellen untersucht. Als Modellpolymere für die Variation der Elastizität wurden vernetzte Poly(n-butylacrylate) (cPnBA) verwendet, da deren Elastizität durch den Anteil des Vernetzers eingestellt werden kann. Auf dem weniger elastischen cPnBA zeigte sich in der Cokultur ein doppelt so hohes Verhältnis von Keratinozyten zu Fibroblasten wie auf dem elastischeren cPnBA, so dass ein leichter zellselektiver Effekt angenommen werden kann. Acrylnitril-basierte Copolymere wurden als Modellpolymere für die Variation der Oberflächenladung und Hydrophilie verwendet, da die Eigenschaften durch Art und molaren Anteil des Comonomers eingestellt werden können. Durch Variation des molaren Anteils der Comonomere mit positiver bzw. negativer Ladung, Methacrylsäure-2-aminoethylester-hydrochhlorid (AEMA) und N-3-Aminopropyl-methacrylamid-hydro-chlorid (APMA) bzw. Natriumsalz der 2-Methyl-2-propen-1-sulfonsäure (NaMAS), wurde der Anteil der positiven bzw. negativen Ladung im Copolymer variiert. Durch die Erhöhung des molaren Anteils des hydrophilen Comonomers N-Vinylpyrrolidon (NVP) wurde die Hydrophilie des Copolymers gesteigert. Die Erhöhung des molaren Anteils an positiv geladenem Comonomer AEMA im Copolymer führte tendenziell zu einer höheren Keratinozytendichte, wobei die Fibroblastendichte unverändert blieb. Durch die Erhöhung des molaren Anteils des positiv geladenen Comonomers APMA ergaben sich keine deutlichen Unterschiede in Dichte, Vitalität oder Selektivität der Zellen. Durch die stufenweise Erhöhung des molaren Anteils des negativ geladenen Comonomers NaMAS konnte, wie im Falle von AEMA, eine Tendenz zur verbesserten Keratinozytenadhärenz beobachtet werden. Die Steigerung der Hydrophilie der Copolymere führte sowohl für Keratinozyten als auch für Fibroblasten zu einer reduzierten Adhärenz und Vitalität. In der vorliegenden Doktorarbeit wurde ein Testverfahren etabliert, das die Untersuchung von primären humanen Keratinozyten und primären humanen Fibroblasten in Monokultur und Cokultur auf verschiedenen Polymeren ermöglicht. Die bisherigen Ergebnisse zeigen, dass sich durch die gezielte Modifizierung verschiedener Polymereigenschaften die Adhärenz und Vitalität beider Zelltypen beeinflussen lässt. Die Reduktion der Elastizität sowie die Erhöhung des molaren Anteils geladener Comonomere führten zu einer Zunahme der Keratinozytenadhärenz. Da die Fibroblasten unbeeinflusst blieben, zeigte sich für einige der untersuchten Polymere eine leichte Zellselektivität. Diese könnte durch die weitere Erhöhung der Steifigkeit oder des Anteils geladener Comonomere möglicherweise weiter gesteigert werden. / Chemical and physical properties of polymers can influence various cell types, e.g. concerning adherence and functionality. For instance, the elasticity of a polymer can influence, which pulling force a cell can generate towards a substrate. According to the cell type, its behavior can be controlled by intracellular feedback mechanisms. The surface charge and/or hydrophilicity of a polymer initially influence the adsorption of ions, proteins and other molecules. In particular, the composition, density, and conformation of the adsorbed components mediate the cell-material interactions. Since different cell types present varying cell membrane associated proteins, sugars and lipids, it is assumed that polymer properties can induce cell specific effects. Polymers, which can regulate specific cell reactions, become more and more important for biotechnological uses and applications in the regenerative medicine. The isolation and culture of primary keratinocytes is still challenging and an adequate wound healing remains a clinical task. A polymer, which enables a preferential adherence of keratinocytes and induces a reduced adherence of dermal fibroblasts, would provide enormous advantages for keratinocyte culture systems as well as for wound dressings. To investigate the specific influence of certain polymer properties on primary human keratinocytes and fibroblasts, a cell culture system for mono- and coculture of both cell types was established. The test system was designed as a screening to investigate the influence of polymers with gradations of different properties on the cells. Thereby, the viability and density of adherent and not adhered cells, as well as the impairment of the cell membranes were analyzed in mono- and cocultures, and the selective adherence of keratinocytes in the coculture was evaluated using a specific immunocytochemical staining for keratin14 and vimentin. Furthermore, the deposition of extracellular matrix components and the secretion of soluble factors were analyzed for the elastic polymers. Since the elasticity of crosslinked poly(n-butylacrylate) (cPnBA) networks can be adjusted by the amount of the crosslinker, they were used as model polymers to investigate the influence of varying elasticity to the cells. On the less elastic cPnBA, the ratio of keratinocytes to fibroblasts was increased compared to the more elastic one. From these results, a slight cell selective effect can be assumed. Acrylonitrile-based copolymers were used as model polymers for the variation of surface charge and hydrophilicity, since their properties can be modified by the type and molar ratio of comonomers. By the variation of the molar ratio of positively charged comonomers (Methacrylic acid-2-aminoethylester hydrochloride (AEMA) and N-3-aminopropyl methacrylamide hydrochloride (APMA)), or a negatively charged comonomer (2-methyl-2-propene-1-sulfonic acid sodium salt (NaMAS)), the amount of positive or negative charges was modified. The hydrophilicity was increased by the molar ratio of the hydrophilic comonomer N-vinylpyrrolidone (NVP). With an increased molar ratio of the positively charged comonomer AEMA, a tendency towards a higher density of adherent keratinocytes could be shown, whereby, the density of adherent fibroblasts remained unaffected. With increasing molar ratios of the positively charged comonomer APMA, no differences between cell densities, viability or selectivity were detectable. Comparable to AEMA, a tendency towards improved keratinocyte adhesion could be shown with an increasing molar ratio of the negatively charged comonomer NaMAS. The increase of the hydrophilicity of the copolymers led to a reduced adherence and viability of the keratinocytes, as well as of the fibroblasts. In conclusion, a test system was established, which enables the evaluation of primary human keratinocytes and fibroblasts in contact with different polymers in monoculture, as well as in coculture. Furthermore, the present thesis shows that directed modifications of polymer properties influenced the adherence and viability of both cell types. The decrease of elasticity and the increase of the molar ratio of charged comonomers led to an increased keratinocyte adherence. Since the fibroblasts remained unaffected, slight cell selectivity was shown. By further increasing the stiffness or the amount of charged comonomers, further enhancement of this effect might be possible.

This dissertation aims to further the understanding of the complex communication that occurs as cells interact with topographical and chemical patterns on a biomaterial interface. The research accomplishes this through two aims fabricating cell substrate surface topography and chemical patterns independently using non-cleanroom approaches, and analyzing higher order cellular response to surface features. The work will impact biomaterial surface modification and fabrication which will apply to biomedical implanted devices, tissue engineering scaffolds, and biological analysis devices. The first aim seeks to apply non-traditional topographical and chemical patterning methods in order to create independent topographical and chemical patterns on cell culture substrates. Experiments use the resulting patterned substrates to quantify cellular alignment to surface topography and compare the relative influence of topographical and chemical patterns on cellular response. The combined patterning methods of imprint lithography and micro-contact printing result in a high-throughput technique applicable to a variety of materials and a range of feature sizes from nanoscale through microscale, thereby enabling future analysis of cell response to surface features. The second aim evaluates the impact of topographical and chemical features on cellular differentiation. Experiments use patterned topography overlaid with a characterized chemical model layer to evaluate the effects of topography on myoblast differentiation and alignment. Chemical patterns that independently control available cell spreading area and modulate cell-cell contact are used to investigate the impact of cell-cell contact on differentiation.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Made available in DSpace on 2015-12-06T23:01:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2001 / O presente estudo teve como objetivo verificar o estresse oxidativo causado a culturas de queratinocitos atraves de sua exposicao a privacao de glicose e a hipoxia, com e sem o uso de um antioxidante, o Dimetil Sulfoxido (DMSO), avaliado atraves da dosagem do malonaldeido (MDA). O material e metodo constituiu-se de tres experimentos. No primeiro deles, 12 garrafas de cultura de queratinocitos foram preparadas ate atingir-se a confluencia desejada e divididas em quatro grupos (com e sem uso de DIVISO, com e sem hipoxia). Foram colhidas amostras do meio de cultura para dosagem do malonaldeido em diferentes fases do experimento: a) pre-experimento, b) 24 horas apos o inicio da privacao de glicose (antes da provocacao de hipoxia), c) 24 horas apos o inicio da privacao de glicose e imediatamente depois da provocacao de hipoxia, d) 48 horas apos a privacao de glicose (24 horas apos a hipoxia). O Experimento 2 constituiu-se das 12 garrafas do final do primeiro experimento e outras 12 garrafas que foram utilizadas como controle, sendo que foi colhido o material celular das 24 garrafas e realizada dosagem do MDA no homogeneizado celular de cada garrafa. O Experimento 3 foi realizado com 80 garrafas divididas em 10 grupos (meio de cultura com e sem glicose, uso ou nao de DIVISO, realizacao de hipoxia ou nao, alem das associacoes entre esses fatores), das quais foi colhido e homogeneizado o material celular para dosagem do MDA. A analise estatistica realizada com os resultados dos tres experimentos mostrou que o DIVISO foi eficiente na reducao do estresse oxidativo de culturas de queratinocitos, causado pela privacao de glicose e hipoxia, avaliado pelos valores de MDA comparando-se aos grupos controle / BV UNIFESP: Teses e dissertações

Submitted by Maria Anália Conceição (marianaliaconceicao@gmail.com) on 2016-06-27T19:07:25Z No. of bitstreams: 1 Publico-NOVO-20.pdf: 888503 bytes, checksum: 0e3c78bc6040c66cc6712fddc9beb7a9 (MD5) / Approved for entry into archive by Maria Anália Conceição (marianaliaconceicao@gmail.com) on 2016-06-27T19:08:15Z (GMT) No. of bitstreams: 1 Publico-NOVO-20.pdf: 888503 bytes, checksum: 0e3c78bc6040c66cc6712fddc9beb7a9 (MD5) / Made available in DSpace on 2016-06-27T19:08:15Z (GMT). No. of bitstreams: 1 Publico-NOVO-20.pdf: 888503 bytes, checksum: 0e3c78bc6040c66cc6712fddc9beb7a9 (MD5) Previous issue date: 2015 / Introdução: Queimadura extensa e profunda é um trauma complexo que necessita de cuidados intensivos no tratamento agudo. Intervenções terapêuticas para atenuar a resposta inflamatória aguda e acelerar a cicatrização podem contribuir na redução da morbimortalidade e nos custos do tratamento. Estudos demonstram a importância do fator de crescimento de queratinócito (KGF) na cicatrização de feridas. Objetivo: Avaliar a ação do KGF na expressão de 84 genes marcadores da cicatrização em cultura primária de queratinócitos humanos oriundos de pacientes com queimadura. Métodos: Após a obtenção de fragmentos de pele de quatro pacientes com queimadura (grupo queimadura) e de quatro pacientes hígidos (grupo controle), foi realizada a cultura de queratinócitos humanos primários e distribuídas em quatro grupos: GQ+ (n = 4 – queratinócitos de queimadura tratadas com KGF), GQ- (n = 4 – queratinócitos de queimadura sem tratamento), GC+ (n = 4 – queratinócitos do grupo controle tratadas com KGF) e GC- (n = 4 – queratinócitos do grupo controle sem tratamento). A análise da expressão gênica foi feita por qPCR Array, realizando seis comparações: 1) GC+ versus GC-; 2) GQ- versus GC-; 3) GQ+ versus GC-; 4) GQ+ versus GQ-, 5) GQ+ versus GC+ e 6) GQ- versus GC+. Resultados: A comparação 1 apresentou um gene hiporregulado e um hiperregulado. As comparações 2 e 3 apresentaram os mesmos cinco genes hiporregulados. A comparação 4 não apresentou genes diferencialmente expressos. A comparação 5 apresentou 26 genes hiporregulados e 7 hiperregulados. E a comparação 6 apresentou 25 genes hiporregulados e 11 genes hiperregulados. Conclusão: A suplementação de KGF à cultura de queratinócitos de pacientes com queimadura não determinou a expressão gênica diferencial dos genes marcadores da cicatrização. / Introduction: Severe burn injury is a complex trauma that needs intensive care in the acute setting. Therapeutic interventions that aim to attenuate the acute inflammatory response and to accelerate the healing may contribute to reducing morbidity, mortality and treatment costs. Several studies have demonstrated the importance of keratinocyte growth factor (KGF) in wound healing. Objective: Evaluate the effect of KGF in the expression of 84 wound healing genes in human primary keratinocytes cultured from patients with burn. Methods: After obtaining viable fragments of skin from four healthy (control group) and four burn patients, human primary keratinocytes was cultured and divided into 4 groups: GQ+ (n = 4 – keratinocytes of severe burn patients treated with KGF), GQ- (n = 4 – untreated keratinocytes of severe burn patients), GC+ (n = 4 – keratinocytes of control group treated with KGF) and GC- (n = 4 – untreated keratinocytes of control group). The gene expression's analysis was performed with qPCR Array, making six comparisons: 1) GC+ versus GC -; 2) GQversus GC-; 3) GQ+ versus GC-; 4) GQ+ versus GQ-, 5) GQ+ versus GC+ and 6) GQversus GC+. Results: Comparison 1 showed one down and one up-regulated genes. Comparisons 2 and 3 showed the same five down-regulated genes. Comparison 4 did not show statistically significant gene expression. Comparison 5 showed 26 downregulated and 7 up-regulated genes. And comparison 6 showed 25 down-regulated and 11 up-regulated genes. Conclusion: Supplementation of KGF to the culture of keratinocytes from patients with burn injury not determined the differential gene expression of genes markers of wound healing

Tissue engineering is an emerging interdisciplinary field developing new ways of treatment of patient's tissue defects using artificial substitutes. Skin tissue engineering is developing skin substitutes and wound dressings that would replace current treatment using autologous, allogeneic or xenogenic substitutes. There are high demands on materials which should serve as a scaffolds for dermal fibroblasts and keratinocytes. They must be non-cytotoxic and biodegradable with a rate proportional to formation of a new tissue. The materials should support adhesion and proliferation of the cells and even they could release growth factors and antimicrobial substance to enhance healing and new tissue formation. In this master thesis, the cell adhesion and proliferation were evaluated on sodium carboxymethyl cellulose (Hcel® NaT), poly-ε-caprolactone (PCL), poly-L-lactide-co-ε-caprolactone (PLA/PCL) and cellulose acetate (AC) nanofiber membranes. Primary human dermal fibroblasts and HaCaT cell line keratinocytes were selected for evaluation. The cell adhesion was observed by fluorescent microscopy, the proliferation was determined by metabolic assay (WST-1) and the material cytotoxicity was evaluated in xCELLigence® system. Materials did not show cytotoxic effects on the cells. However, the materials did...

La ciguatéra est une forme d’intoxication faisant suite à l’ingestion de poissons contaminés par des toxines appelées « ciguatoxines ». Cette intoxication endémique des régions tropicales est un problème économique et de santé non négligeable qui tend à prendre de plus en plus d’ampleur. L’essor du tourisme, le réchauffement climatique et la hausse des exportations internationales de poissons tropicaux favorisent l’expansion de la ciguatéra aux parties du globe au climat tempéré, jusqu’alors peu concernées par cette maladie. Les enjeux thérapeutiques et économiques de la ciguatéra sont de taille puisqu’il n’existe aucun moyen rapide et fiable de détecter un poisson contaminé et qu’aucun traitement efficace permettant sa prise en charge n’a actuellement été établi.Le prurit, terme médical désignant les démangeaisons est un symptôme notamment associé aux maladies de peau qui impacte grandement la qualité de vie des patients qui en souffrent.Ces dernières décennies, de nombreuses études scientifiques ont permis de mieux comprendre sa physiopathologie. Le prurit est un symptôme fréquemment observé chez les personnes atteintes de ciguatéra, d’où l’appellation « la Gratte », souvent employée pour faire référence à la pathologie.Dans le but d’étudier les mécanismes cellulaires à l’origine du prurit et des autres troubles sensoriels cutanés survenant lors de la ciguatéra, nous avons étudié l’effet des ciguatoxines sur un modèle in vitro de neurones sensoriels cocultivés avec des kératinocytes. Nous avons mis en évidence la libération dans le surnageant des neuropeptides de l'inflammation neurogène, SP et CGRP. L'effet d'antagonistes sélectionnés a été testé afin mettre en évidence les médiateurs impliqués dans la libération de neuropeptides. Par ailleurs, la signalisation cellulaire sous-jacente de cette sécrétion de neuropeptides a été étudiée par des expériences d’imagerie calcique réalisées sur neurones et kératinocytes.Les résultats obtenus valident notre coculture en tant que modèle de choix pour l’étude in vitro des mécanismes cellulaires, et plus généralement, dans les troubles neurocutanés impliqués dans le prurit ciguatérique. Parmi les antagonistes testés, une molécule s’est avérée particulièrement intéressante pour inhiber les effets constatés de la toxine. Ces résultats originaux, obtenus avec l’antagoniste d’un médiateur du prurit, présentent une perspective thérapeutique nouvelle et prometteuse, pour répondre à un enjeu de santé publique futur. / Ciguatera fish poisoning (CFP) is a seafood poisoning occurring after contaminated fish fleshes ingestion containing toxins called « ciguatoxines » (CTXs). This illness, originating from tropical and subtropical areas, is an economic and health problems which becomes substantial in relation to international tropical fishes export and tourism development, as well as global warming rise. Those factors contribute to CFP sprouting in non-endemic temperate climate regions which were not until then concerned by CFP. Economic and health stakes of CFP are important since no reliable and ready-to-use detection system for CTXs in fishes have been developed, along with no relevant cure has been established to treat CFP.Pruritus, medical term refer to itch, is a clinical sign usually associated to skin diseases which strongly alter patients’ quality of life. Last decades, several studies allowed to better understand pruritus pathophysiology. Interestingly, people suffering from CFP frequently present pruritus, hence designation “La Gratte” or “La Gratel (le)” employed in endemic areas.The aim of these works was to study cellular and molecular mechanisms of CTXs at the root of neurological cutaneous troubles occurring in CFP. Here, we evaluated CTXs effects on in vitro model composed of sensory neurons cocultived with primary keratinocytes, quantifying neuropeptides known to be involved in pruritus. Compiling knowledges about CFP and pruritus pathophysiology, some antagonists were tested to neutralize CTXs-mediated neuropeptide release. To deal with signaling pathways in depth of neuropeptide exocytosis induced by CTXs, mechanism known to be accurately regulated by calcium homeostasis, calcium imaging experiments were performed.Results obtained in this project confirm the use of such a model to elucidate cellular mechanism of CFP pruritus, but also constitute an alternative in vitro tool to study chemicals inducing abnormal cutaneous senses. Among antagonists tested, one stands out from the crowd and was proved to be effective to inhibit CTXs-evoked effects studied. Those originals results, collected with antagonist of pruritus mediator, show new and promiscuous therapeutic prospects for future health concern.