Der Transkriptionsfaktor NF-κB ist von großer Bedeutung, da er verschiedene zelluläre Prozesse wie Proliferation, Apoptose, Invasion oder Inflammation beeinflusst. Im Gegensatz zu den meisten anderen Zelltypen ist in der humanen Epidermis ein wachstumsinhibierender Effekt mit der Aktivierung von NF-ĸB assoziiert. Die epidermale Homöostase dient der Aufrechterhaltung der intakten Hautbarriere und beschreibt das Gleichgewicht zwischen proliferierenden und differenzierenden epidermalen Keratinozyten. Schädliche äußere Einflüsse wie übermäßige Sonnenlichtexposition können die epidermale Homöostase stören, was zur Entstehung epidermaler Neoplasien wie aktinischer Keratosen oder Plattenepithelkarzinomen beiträgt. Das Ziel dieser Arbeit war die Expressions- und Funktionsanalyse der einzelnen NF-κB Proteinuntereinheiten in humanen Keratinozyten in vitro. Die siRNA-vermittelte transiente Reduktion von c-Rel beeinflusste deutlich das Zellschicksal von Keratinozyten. Obwohl vorangegangene Experimente durch eine stärkere Zellproliferation nach Inhibierung der NF-ĸB Proteine p50 und p65 das Gegenteil suggerierten, konnte für die Reduktion von c Rel ein inhibierender Effekt auf das Zellwachstum festgestellt werden. Außerdem zeigten sich eine veränderte Zellzyklusphasenverteilung sowie eine Akkumulation mitotischer Zellen mit aberranter, hauptsächlich monopolarer Spindelformation. Die zusätzlich detektierte Apoptose-Induktion könnte aus dem verlängerten mitotischen Arrest der c-Rel Knockdown Keratinozyten resultieren. Insgesamt lässt sich ein regulatorischer Effekt von c-Rel beim Eintritt in die Mitose oder der mitotischen Progression vermuten, wobei die beteiligten Zielgene noch zu identifizieren sind. Des Weiteren bewirkte der c-Rel Knockdown phänotypische Veränderung von HaCaT Keratinozyten mit tendenziell spindelzellartiger Elongation und einem insgesamt veränderten Wachstumsmuster. Die Adhäsion und besonders die Wundheilung von c-Rel reduzierten HaCaT Zellen war vermindert, möglicherweise bedingt durch ein reduziertes Stressfaservorkommen. Dieser Effekt zeigte sich allerdings nicht in c Rel herunter regulierten primären Keratinozyten, was auf Mutationen der spontan immortalisierten HaCaT Keratinozytenzelllinie zurückzuführen sein könnte. Zusammenfassend konnte in dieser Arbeit ein neuer Aspekt der einzelnen NF-ĸB Proteine aufgezeigt werden, besonders in Bezug auf die Proteinuntereinheit c-Rel. Hieraus resultiert ein besseres Verständnis der vielfältigen und komplexen Regulation von NF-κB abhängigen Funktionen und deren Effekte auf die epidermale Homöostase.

No description available.

no / Eczema is widely considered to be an exacerbation of alkaline stress to the skin. Epidermal barrier dysfunction is a feature of eczema pathology, which predisposes affected individuals to distressing morbid symptoms. At least two serine proteases, stratum corneum chymotryptic enzyme (kallikrein 7 [KLK7]) and stratum corneum tryptic enzyme (kallikrien 5 [KLK5]), have increased activity levels in eczematous lesions and both have been implicated in the destruction of corneodesomosomes, which are crucial to epidermal integrity. The present in vitro study investigated whether transcriptional gene silencing after siRNA transfection could influence the activity of these signature enzymes in an in vitro model of eczema induced by alkaline shock. HaCaT epithelial cells were subjected to alkaline stress by the addition of 1,1,3,3-tetramethyl guanidine “superbase” (TMG) to the culture media. The culture media were subsequently tested for chymotryspin, trypsin, plasmin, and urokinase activity using colorimetric peptide assays and for reactive oxygen species using WST1 cell viability reagent. Cells that had been transfected with small interfering ribonucleic acid (siRNA) against KLK5 and KLK7 for 24 h before alkaline shock did not exhibit the increase in serine protease levels observed in untreated controls. Moreover, an endpoint MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) confirmed that detachment of cells from the culture substrate observed in alkaline-stressed cells did not occur in siRNA-treated cells. This in vitro study has established the proof-of-principle that siRNA therapy appears to mitigate the consequences of alkaline shock to the serine protease-associated fragility of epithelial cells that is characteristic of eczema.

Nanotechnology applications have emerged enormously in recent times. Of particular interest is that area that overlaps the areas of nanotechnology, biology and medicine: nanomedicine. One advantage of nanomedicines is it that it can be used as an enabling technology by pharmaceutical researchers and industry to overcome issues associated with the low bioavailability of hydrophobic drugs. In the first part of the current study, nanosuspensions of two of hydrophobic steroid drugs: hydrocortisone and prednisolone were produced. Nanosuspensions were prepared using a bottom-up approach: the anti-solvent precipitation method using microfluidic reactors. Surface modification was carried out on these nanosuspensions using cationic surfactants to obtain nanoparticles with different levels of surface positive charge as indicated by ¿-potential values. Dynamic light scattering (DLS) and transmission electron microscope (TEM) techniques were used to characterize the prepared nanoparticles. Powder X-ray diffraction (PXRD) and differential scanning calorimetry (DSC) were also used to characterize hydrocortisone nanoparticles. In the second part, cellular uptake of both coated and uncoated nanoparticles by HaCaT keratinocytes cell line was examined and indicated by quantifying the anti- inflammatory effect of nanoparticles on the LPS-induced inflammation. Also, TEM was employed to evaluate the cellular uptake of hydrocortisone nanoparticles. Results showed higher ant-inflammatory effect of coated nanoparticles over uncoated nanoparticles. Furthermore, the anti-inflammatory effect of coated nanoparticles was correlated to the degree of positive surface charge. / Libyan government

No / The use of nanoparticles have provided numerous of advantages in medicine due to their unique physiochemical characteristics such as size, charge, shape and surface reactivity [1-4]. Understanding the interaction between engineered nanomaterials and living matter has attracted increasing attention in recent years. Toxicity of nanoparticles was studied in different cell types and cell lines. Nano-SiO2 has good stability, easy dispensability, and melting degeneration, and is widely used in rubber, paints, biomedical and biotechnology fields [5]. In this study, the LDH assay and the MTT assay were applied to evaluate the cytotoxicity of in vitro Silicon dioxide nanoparticles (SiO2-NP, 20nm) on cultured cell lines. Human lung adenocarcinoma epithelial cell line (A549) were used as a lung related cell line and human keratinocyte cell line (HaCaT) as a skin related cell line representing different uptake routes. The percentage cytotoxicity of the silicon dioxide nanoparticles was measured once cultured in a 24 hour incubation period. The concentration of the SiO2 nanoparticles chosen was 10, 50, 100 and 200µg/ml. To measure the cytotoxicity of nanoparticle on cultured cell lines, we used 104*cells/100 µl of cell culture media being placed in a 96 well rounded bottom plate with the LDH assay. The extracellular lactate dehydrogenase release was measured by using a colorimetric CytoTox 96 non-radioactive assay kit and the absorbance was recorded at 492nm. The MTT assay was used to evaluate mitochondrial activity which includes cell growth and cell death. This has been performed by inserting a premixed optimized dye solution in the culture wells. The Absorbance was recorded at 570 nm, from the recorded absorbance is directly proportional to the number of live cells. In order to maintain the cell lines, they were placed in a plastic T-75cm² tissue culture flasks grown in Dulbecco's Modified Eagle's Medium. Studies were performed in the absence of serum. Cytotoxicity was found in both cells the A549 and HaCaT cells and cytotoxicity increased as concentration of the silicon dioxide increased. The percentage cytotoxicity calculated was higher in HaCaT cells compared to the A549 cells. A cell count assay was plated in order to display the cell number of both the HaCaT and A549 cells. The cell count reaffirmed that cytotoxicity did occur as the cell count decreased as the concentration of the silicon dioxide increased compared to the control. These results show that silicon dioxide nanoparticles acted differently in two different cell types and that the metabolic rate of a cell can be used to determine the nanoparticles affect. Further understanding of the mechanism involving the ROS generation could provide more information on how silicon dioxide nanoparticles increase cytotoxicity. / Physiology 2015 conference abstract

Leaves from Combretum collinum Fresen (Combretaceae) are commonly used for the treatment of inflammatory conditions, wound healing and bacterial infections in traditional West African medicine. This research focuses on the characterization of the phenolic profile and lipophilic compounds of leaves extracts of C. collinum. Studies of the in vitro anti-inflammatory activity were performed in TNFα stimulated HaCaT cells and antibacterial activity was evaluated with agar well diffusion and microdilution assays. Antioxidant activity was determined by DPPH and ABTS assays and compared to standards. The phytochemical studies confirmed myricetin-3-O-rhamnoside and myricetin-3-O-glucoside as major components of the leaves extracts, each contributing significantly to the antioxidant activity of the hydrophilic extracts. GC-MS analysis identified 19 substances that were confirmed by comparison with spectral library data and authentic standards. Combretum collinum aqueous leaves extract decreased pro-inflammatory mediators in TNFα stimulated HaCaT cells. Further investigations showed that myricetin-3-O-rhamnoside has an anti-inflammatory effect on IL-8 secretion. In the antimicrobial screening, the largest inhibition zones were found against S. epidermidis, MRSA and S. aureus. MIC values resulted in 275.0 µg/mL for S. epidermidis and 385.5 µg/mL for MRSA. The in vitro anti-inflammatory, antibacterial and antioxidant activity supports topical use of C. collinum leaves extracts in traditional West African medicine.

yes / Ibuprofen (IBF) is an outstanding non-steroidal drug for analgesic and anti-inflammatory therapies but it exhibits poor solubility in water [1, 2]. Increased dosage administration has been linked to gastrointestinal and cardiovascular complications [3]. Many techniques have been employed to improve the solubility of NSAIDs [4]. In this study, the anti-solvent precipitation method was used to make Ibuprofen nanoparticles (IBF NPs). Optimised preparation parameters such as solvent (ethanol), raw drug concentration (400 mg), solvent/anti-solvent ratio (1:50) and surfactant concentration (0.25 mg/ml) have been studied to yield nanoparticles with a mean size of 58.8 nm, which is confirmed by dynamic light scattering and transmission electron microscopy. These IBF NPs posess increased aqueous solubility compared to the micro counterpart and maintain with chemical integrity indicated by high performance liquid chromatography and Fourier transform infrared spectroscopy. In addition, in vitro cytotoxicity of IBF NPs has been studied on A549 and HaCat cell lines using MTT and LDH assays. Both cells were obtained from ATCC. The A549 cells were grown in a modification of Ham’s F-12, containing L-glutamine, called F-12K. The HaCaT cells were grown in DMEM containing sodium pyruvate (110 mg/l). Normal cell culture and sub-culture were applied and the cells were used after around 45 passages [5]. The cell culture media containing 105cells/ml were placed in a 96-well plate with addition of IBF NPs and Micro form at concentrations in the range of between 6 and 500 ug/ml by diluting them with DMEM and F-12K for use with the HaCaT and A549 cells respectively. After 24, 48 and 72h exposure, the MTT and LDH cytotoxicity assay were performed in triplicates and on three separate experiment cultures and the absorbance was recorded at 570 nm and 492nm respectively with Elisa micro plate reader. The cell viability (%) related to control (cells in culture medium without NPs) was calculated. A very good cytotoxicity profile was observed, indicating an in vitro cytocompatibility of the IBF NPs in these cell culture systems and no significant changes in cytotoxicity compared with Micro IBF. We conclude that our IBF NPs have increased solubility, same chemical integrity and unchanged cytotoxicity compared to IBF Micro drug. Further work will concentrate on optimising more rigorous parameter to produce excellent quality NPs. More detailed characterisation of IBF NPs is to be tested, such as using PXRD and SEM to further corroborate particle shape and size. The range of no toxic in vitro concentrations is also to be further confirmed. Eventually scaled up preparation of IBF NPs will be developed without relinquishing NPs quality. This would improve the potential for in vitro/ in vivo applications and clinical use of IBF NPs and NSAIDs in general.

yes / Nanoparticles are increasingly used in industry and medicine due to their unique physiochemical properties such as their small size, charge, shape, chemical architecture, large surface area, surface reactivity and media interactions, etc [1-5]. However, very little is still known on the interactions between nanoparticles and the biological system. This study aims to evaluate the cytotoxicity of polystyrene latex bead nanoparticles on HaCat and HeLa cell lines. Carboxyl-modified 20 nm polystyrene NPs core labelled with fluorophore were from Invitogen. We chose to use polystyrene NPs because this specific type of NP is being increasingly characterized for use in nanosensors and drug nanocarrier investigations. 1x 104 cells/100 μl of cell culture medium were plated into 96-well plates in triplicate, measuring activity post 24 hours at concentrations of 10, 50, 100 μg/ml of polystyrene NPs exposure. The extracellular lactate dehydrogenase release was measured by using a colorimetric CytoTox 96 nonradioactive assay kit from Promega and the absorbance were recorded at 450nm (FLUO-star) with Elisa micro plate reader. The MTT assay was used to evaluate mitochondrial activity. This was performed by inserting a premixed optimized dye solution in the culture wells. The Absorbance was recorded at 570 nm, from the recorded absorbance is directly proportional to the number of live cells. The cell lines were kept in a plastic T-75cm2 tissue culture flasks grown in DMEM. We found that cytotoxicity of polystyrene NPs on both cells was concentration dependent. For Hela cells, with exporesure of polystyrene NPs at concentrations of 10, 50, 100 μg/ml for 24 hrs, the percentage cytotoxicity of positive control for LDH assay was 35.9%, 49.5% and 73.4% respectively. With the MTT cell viability assay the percentage MTT reduction of negative control was 88.9%, 42.9% and 26.4% respectively. Cell toxicity increased with increasing polystyrene NPs concentration. For HaCaT cells, the cytotoxic effect is less significant than those on Hela cells. With MTT assay, when compared to HaCaT cells exposed to a negative control containing only PBS, the cell viability decreased as the concentrations of NPs increased. Cells exposed to 100μg/ml of polystyrene NPs for a period of 24 hours compared to those exposed to a positive control (100% cell viability) had an average cell viability of 49%, with those numbers decreasing from 59% for cells exposed to 10μg/ml of polystyrene NPs to 57% for cells exposed to 50μg/ml of polystyrene NPs. Our results indicated that polystyrene NPs acted differently in two different cell types and that cautions should be taken about its cytotoxicity. Further understanding of the mechanism involving the ROS generation could provide more information on how polystyrene NPs increase cytotoxicity.

O tingimento de tecidos começou há milhares de anos e a disponibilidade comercial de corantes é enorme e crescente. A indústria têxtil brasileira desempenha um papel de inquestionável importância, destacando-se entre as principais atividades econômicas do país. O processo de tingimento é um dos fatores fundamentais no sucesso comercial dos produtos têxteis, uma vez que o consumidor exige cores resistentes à exposição ao calor, à luz, à transpiração e às lavagens. Segundo a literatura, condições de transpiração intensa contribuem para uma alta taxa de migração e subseqüente penetração de corantes têxteis para a pele humana. Além disso, 2 a 50% desses compostos permanecem no banho de tingimento e são descartados nos efluentes industriais, contaminando o ambiente e colocando em risco a saúde humana, uma vez que os métodos convencionais de tratamento de efluentes são ineficientes na remoção da coloração e da mutagenicidade de alguns corantes. Dentro deste contexto, este trabalho teve como objetivo avaliar os efeitos toxicogenéticos do corante Direct Black (DB38) original e após extração por lixiviação com suor sintético, utilizando o teste do cometa com fibroblastos e queratinócitos de pele humana, o teste Anexina V com fibroblastos e o ensaio de mutagenicidade com Salmonella typhimurium. Adicionalmente, foi investigada a ecotoxicidade dos corantes têxteis Direct Black 38 e Reactive Blue 15 (RB15) originais por meio de ensaios com sementes, dapnhias, minhocas e zebrafish realizados na UTOX, em Barcelona. O corante DB38 original e lixiviado não induziram genotoxicidade em fibroblastos e queratinócitos de pele humana. O corante DB38 original foi mutagênico para as linhagens TA98 e TA100 de S. typhimurium na presença de S9. Entretanto, o corante lixiviado não induziu mutagenicidade para essas linhagens testadas, considerando que a maior taxa de migração do corante para a solução de suor foi de ~1% nas seguintes condições: tingimento sem ensaboamento, pH 8,0 e 8 horas de incubação à 42°C. O corante original é citotóxico para fibroblastos após 48 horas de exposição. No entanto, essa citotoxicidade não foi mais observada após a lixiviação no suor. Os corantes DB38 e RB15 originais não foram tóxicos para as sementes de pepino, alface e tomate, e nem para as minhocas Eisenia foetida. Ambos os corantes foram fracamente tóxicos para Daphnia magna, porém o RB15 apresenta maior potencial tóxico em relação ao DB38. Os corantes DB38 e RB15 induziram malformações em larvas de zebrafish Danio rerio, caracterizadas por falha na inflação da bexiga natatória e alteração na cauda. Portanto, nossos resultados mostram a importância de se fazer não só a análise individual de corantes têxteis, mas também dos tecidos que os contêm. Além da necessidade de se desenvolver técnicas de tingimento mais seguras em relação à solidez da cor sob condições úmidas e as perdas de corante para o ambiente durante a etapa de fixação, indicando maior atenção ao estudo de efeitos sub-letais na avaliação do impacto desses compostos no ecossistema aquático. / The fabrics dyeing began thousands of years ago and the commercial availability of dyes is increasingly. The Brazilian textile industry plays a role of high importance, highlighting among the main economic activities in the country. The dyeing process is one of the key factors in the commercial success of textile products, since consumers are demanding colors more resistant to heat, light exposure, perspiration and washing. According to the literature, conditions of intense perspiration contribute to the migration and subsequent penetration of textile dyes to human skin. Furthermore, 2 to 50% of the initial dye load is present in the dye bath effluent and these compounds are discharged in industrial effluents, contaminating the environment and endangering human health, since the wastewater treatment systems are ineffective in removing the color and mutagenicity of some dyes. In this context, this study aimed to evaluate the toxicogenetic effects of the Direct Black 38 (DB38) dye original and extracted by leaching with artificial sweat using Comet assay with fibroblasts and keratinocytes from human skin, Anexin V assay with fibroblasts and Salmonella mutagenicity test. Additionally, we investigated the ecotoxicity of textile dye Direct Black 38 and Reactive Blue 15 (RB15) using assays with seeds, dapnhias, worms and zebrafish performed in UTOX in Barcelona. The original and leached DB38 dye did not induce genotoxicity in fibroblasts and keratinocytes from human skin. The original DB38 was mutagenic for TA98 and TA100 of S. typhimurium with S9. However, the solution with the leached dye did not induce mutagenicity for these tested strains, since the highest migration rate of the dye to the solution of artificial sweat was ~ 1% in the following conditions: type of dyeing without rinsing, pH 8.0 and 8-hour incubation at 42°C. The original dye was cytotoxic for fibroblasts after 48 hours of exposure. However, this cytotoxicity was no longer observed after leaching in sweat. The original DB38 and RB15 dyes showed no toxicity for cucumber, lettuce and tomato seeds and for earthworms Eisenia foetida. Both dyes were weakly toxic for Daphnia magna, but the RB15 has a higher toxic potential compared to DB38. The dyes DB38 and RB15 induced malformations in larvae of zebrafish Danio rerio by failure of the swim bladder inflation and changes in the tail. Therefore, our results show the importance of making the individual analysis of textile dyes, but also of fabrics containing them. Furthermore, it is necessary to develop safer techniques of dyeing in relation to the color fastness under humid conditions and the loss of dyes into the environment during the fixation step, indicating more attention to the study of sub-lethal effects in the evaluation of the impact of these compounds in the aquatic ecosystem.

O tingimento de tecidos começou há milhares de anos e a disponibilidade comercial de corantes é enorme e crescente. A indústria têxtil brasileira desempenha um papel de inquestionável importância, destacando-se entre as principais atividades econômicas do país. O processo de tingimento é um dos fatores fundamentais no sucesso comercial dos produtos têxteis, uma vez que o consumidor exige cores resistentes à exposição ao calor, à luz, à transpiração e às lavagens. Segundo a literatura, condições de transpiração intensa contribuem para uma alta taxa de migração e subseqüente penetração de corantes têxteis para a pele humana. Além disso, 2 a 50% desses compostos permanecem no banho de tingimento e são descartados nos efluentes industriais, contaminando o ambiente e colocando em risco a saúde humana, uma vez que os métodos convencionais de tratamento de efluentes são ineficientes na remoção da coloração e da mutagenicidade de alguns corantes. Dentro deste contexto, este trabalho teve como objetivo avaliar os efeitos toxicogenéticos do corante Direct Black (DB38) original e após extração por lixiviação com suor sintético, utilizando o teste do cometa com fibroblastos e queratinócitos de pele humana, o teste Anexina V com fibroblastos e o ensaio de mutagenicidade com Salmonella typhimurium. Adicionalmente, foi investigada a ecotoxicidade dos corantes têxteis Direct Black 38 e Reactive Blue 15 (RB15) originais por meio de ensaios com sementes, dapnhias, minhocas e zebrafish realizados na UTOX, em Barcelona. O corante DB38 original e lixiviado não induziram genotoxicidade em fibroblastos e queratinócitos de pele humana. O corante DB38 original foi mutagênico para as linhagens TA98 e TA100 de S. typhimurium na presença de S9. Entretanto, o corante lixiviado não induziu mutagenicidade para essas linhagens testadas, considerando que a maior taxa de migração do corante para a solução de suor foi de ~1% nas seguintes condições: tingimento sem ensaboamento, pH 8,0 e 8 horas de incubação à 42°C. O corante original é citotóxico para fibroblastos após 48 horas de exposição. No entanto, essa citotoxicidade não foi mais observada após a lixiviação no suor. Os corantes DB38 e RB15 originais não foram tóxicos para as sementes de pepino, alface e tomate, e nem para as minhocas Eisenia foetida. Ambos os corantes foram fracamente tóxicos para Daphnia magna, porém o RB15 apresenta maior potencial tóxico em relação ao DB38. Os corantes DB38 e RB15 induziram malformações em larvas de zebrafish Danio rerio, caracterizadas por falha na inflação da bexiga natatória e alteração na cauda. Portanto, nossos resultados mostram a importância de se fazer não só a análise individual de corantes têxteis, mas também dos tecidos que os contêm. Além da necessidade de se desenvolver técnicas de tingimento mais seguras em relação à solidez da cor sob condições úmidas e as perdas de corante para o ambiente durante a etapa de fixação, indicando maior atenção ao estudo de efeitos sub-letais na avaliação do impacto desses compostos no ecossistema aquático. / The fabrics dyeing began thousands of years ago and the commercial availability of dyes is increasingly. The Brazilian textile industry plays a role of high importance, highlighting among the main economic activities in the country. The dyeing process is one of the key factors in the commercial success of textile products, since consumers are demanding colors more resistant to heat, light exposure, perspiration and washing. According to the literature, conditions of intense perspiration contribute to the migration and subsequent penetration of textile dyes to human skin. Furthermore, 2 to 50% of the initial dye load is present in the dye bath effluent and these compounds are discharged in industrial effluents, contaminating the environment and endangering human health, since the wastewater treatment systems are ineffective in removing the color and mutagenicity of some dyes. In this context, this study aimed to evaluate the toxicogenetic effects of the Direct Black 38 (DB38) dye original and extracted by leaching with artificial sweat using Comet assay with fibroblasts and keratinocytes from human skin, Anexin V assay with fibroblasts and Salmonella mutagenicity test. Additionally, we investigated the ecotoxicity of textile dye Direct Black 38 and Reactive Blue 15 (RB15) using assays with seeds, dapnhias, worms and zebrafish performed in UTOX in Barcelona. The original and leached DB38 dye did not induce genotoxicity in fibroblasts and keratinocytes from human skin. The original DB38 was mutagenic for TA98 and TA100 of S. typhimurium with S9. However, the solution with the leached dye did not induce mutagenicity for these tested strains, since the highest migration rate of the dye to the solution of artificial sweat was ~ 1% in the following conditions: type of dyeing without rinsing, pH 8.0 and 8-hour incubation at 42°C. The original dye was cytotoxic for fibroblasts after 48 hours of exposure. However, this cytotoxicity was no longer observed after leaching in sweat. The original DB38 and RB15 dyes showed no toxicity for cucumber, lettuce and tomato seeds and for earthworms Eisenia foetida. Both dyes were weakly toxic for Daphnia magna, but the RB15 has a higher toxic potential compared to DB38. The dyes DB38 and RB15 induced malformations in larvae of zebrafish Danio rerio by failure of the swim bladder inflation and changes in the tail. Therefore, our results show the importance of making the individual analysis of textile dyes, but also of fabrics containing them. Furthermore, it is necessary to develop safer techniques of dyeing in relation to the color fastness under humid conditions and the loss of dyes into the environment during the fixation step, indicating more attention to the study of sub-lethal effects in the evaluation of the impact of these compounds in the aquatic ecosystem.