Triclosan and Triclocarban, preservatives widely used in cosmetics and other consumer products, underwent evaluation using a battery of new-approach methodologies in vitro (NAMs). Specifically, the Microplate Ames Test (MPFTM Test, Xenometrix, Allschwil, Switzerland) was employed to assess mutagenicity, the Comet assay in vitro on the HaCat cell line and the Mammalian Chromosome Aberration Test were utilized to evaluate genotoxicity, and the XenoScreen® YES/YAS assay was applied to investigate endocrine disruption. The chemicals did not exhibit any positive responses for mutagenicity. However, the mammalian chromosome aberration test identified both chemicals as being positive for genotoxicity at 10 μg/mL. In the Comet assay, the percentage of DNA in the tail significantly increased in a concentration-dependent manner (at 5 and 10 μg/mL for Triclosan, at 2.5, 5, and 10 μg/mL for Triclocarban). The positive response depended on the increasing concentration and the duration of exposure. Triclosan, but not Triclocarban in any of the endocrine assays performed, indicated a potential for endocrine activity in the anti-estrogenic and anti-androgenic assays. The positive in vitro results detected were obtained for concentrations relevant to final products. The alarming findings obtained with the use of new-approach methodologies (NAMs) justify the current precautionary regulatory approach, limiting the use of these preservatives.
- Publikační typ
- časopisecké články MeSH
Animal testing has been prohibited for the safety assessment of cosmetic ingredients or finished products. Thus, alternative non-animal methods, followed by confirmatory clinical studies on human volunteers, should be used as the sole legally acceptable approach within the EU. The safety assessment of cosmetic products requires the involvement of multiple scientific disciplines, including analytical chemistry and biomedicine, as well as in chemico, in vitro and in silico toxicology. Recent data suggest that fragrance components may exert multiple adverse biological effects, e.g. cytotoxicity, skin sensitisation, (photo)genotoxicity, mutagenicity, reprotoxicity and endocrine disruption. Therefore, a pilot study was conducted with selected samples of fragrance-based products, such as deodorant, eau de toilette and eau de parfum, with the aim of integrating results from a number of alternative non-animal methods suitable for the detection of the following toxicological endpoints: cytotoxicity (with 3T3 Balb/c fibroblasts); skin sensitisation potential (in chemico method, DPRA); skin sensitisation potential (LuSens in vitro method, based on human keratinocytes); genotoxicity potential (in vitro Comet assay with 3T3 Balb/c cells); and endocrine disruption (in vitro YES/YAS assay). The presence of twenty-four specific known allergens in the products was determined by using GC-MS/MS. The strategies for estimation of the NOAEL of a mixture of allergens, which were proposed by the Scientific Committee on Consumer Products in their 'Opinion on Tea tree oil' document and by the Norwegian Food Safety Authority in their 'Risk Profile of Tea tree oil' report, were used as models for the NOAEL estimation of the mixtures of allergens that were identified in the individual samples tested in this study.
- MeSH
- alergeny toxicita analýza MeSH
- kosmetické přípravky * toxicita MeSH
- lidé MeSH
- parfém * analýza MeSH
- pilotní projekty MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- tandemová hmotnostní spektrometrie MeSH
- tea tree oil * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVE: Precise control over the ultrasound field parameters experienced by biological samples during sonication experiments in vitro may be quite challenging. The main goal of this work was to outline an approach to construction of sonication test cells that would minimize the interaction between the test cells and ultrasound. METHODS: Optimal dimensions of the test cell were determined through measurements conducted in a water sonication tank using 3D-printed test objects. The offset of local acoustic intensity variability inside the sonication test cell was set to value of ±50% of the reference value (i.e., local acoustic intensity measured at last axial maximum in the free-field condition). The cytotoxicity of several materials used for 3D printing was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. RESULTS: The sonication test cells were 3D printed from polylactic acid material, which was not toxic to the cells. Silicone membrane HT-6240, which was used to construct the bottom of the test cell, was found to reduce ultrasound energy minimally. Final ultrasound profiles inside the sonication test cells indicated the desired variability of local acoustic intensity. The cell viability in our sonication test cell was comparable to that of commercial culture plates with bottoms constructed with silicone membrane. CONCLUSION: An approach to construction of sonication test cells minimizing the interaction of the test cell and ultrasound has been outlined.
Photodynamic therapy is an alternative treatment mainly for cancer but also for bacterial infections. This treatment dates back to 1900 when a German medical school graduate Oscar Raab found a photodynamic effect while doing research for his doctoral dissertation with Professor Hermann von Tappeiner. Unexpectedly, Raab revealed that the toxicity of acridine on paramecium depends on the intensity of light in his laboratory. Photodynamic therapy is therefore based on the administration of a photosensitizer with subsequent light irradiation within the absorption maxima of this substance followed by reactive oxygen species formation and finally cell death. Although this treatment is not a novelty, there is an endeavor for various modifications to the therapy. For example, selectivity and efficiency of the photosensitizer, as well as irradiation with various types of light sources are still being modified to improve final results of the photodynamic therapy. The main aim of this review is to summarize anticancer and antibacterial modifications, namely various compounds, approaches, and techniques, to enhance the effectiveness of photodynamic therapy.
- MeSH
- antibakteriální látky farmakologie terapeutické užití MeSH
- buněčná smrt MeSH
- fotochemoterapie * metody MeSH
- fotosenzibilizující látky farmakologie terapeutické užití MeSH
- lidé MeSH
- nádory * farmakoterapie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Growing worldwide efforts to replace (reduce) animal testing and to improve alternative in vitro tests which may be more efficient in terms of both time, cost and scientific validity include also genotoxicity/mutagenicity endpoints. The aim of the review article was to summarize currently available in vitro testing approaches in this field, their regulatory acceptance and recommended combinations for classification of chemicals. A study using the combination of Comet Assay performed on two cell lines and the Chromosomal Aberration test on human peripheral lymphocytes was performed with the aim to predict the genotoxic potential of selected paraben esters, serving as a model chemical group. Parabens are widely used in consumer products as preservatives and have been reported to exhibit inconclusive results in numerous genotoxicity studies. The Comet Assay identified Ethylparaben and Benzylparaben as potentially genotoxic. The Chromosomal Aberration test revealed weak genotoxic potential in case of Ethylparaben and positive genotoxicity in case of Butylparaben, Propylparaben and Isopropylparaben. The main reasons for variability seem to be limited water solubility of parabens, determining their bioavailability at the cellular level, and absence of metabolic activation in the Comet Assay. The results confirmed that the Comet Assay should serve as a screening test and should not be used as a stand-alone method for classification of genotoxicity. The weight of evidence approach in risk assessment should be supported with data generated with the use of human relevant in vitro methods based on cells / tissues of human origin.
- MeSH
- alternativy testů na zvířatech * MeSH
- buněčné linie keratinocytů HaCaT MeSH
- chromozomální aberace chemicky indukované MeSH
- hodnocení rizik MeSH
- kometový test MeSH
- lidé MeSH
- lymfocyty účinky léků patologie MeSH
- mikrojaderné testy MeSH
- mikrojádra chromozomálně defektní chemicky indukované MeSH
- mutageneze účinky léků MeSH
- parabeny toxicita MeSH
- poškození DNA * MeSH
- testy genotoxicity * MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- srovnávací studie MeSH
- Publikační typ
- abstrakt z konference MeSH
Rose bengal and erythrosin B are xanthene dyes mainly known and used as antimicrobial agents, but due to their photodynamic activity they are also potential photosensitizers for cancer photodynamic therapy. The aim of this work is to study a photodynamic efficacy of rose bengal and erythrosin B against human skin melanoma and mouse fibroblast cell lines, compare them with each other and find out their photodynamic properties induced by light emitting diodes with total light dose of 5 J/cm2. To fully identify and understand photodynamic properties of both potentially effective photo-sensitizers, a set of complex in vitro tests such as cell cytotoxic assay, measurement of reactive oxygen species production, mitochondrial membrane potential change assay, mode of cell death determination or comet assay were made. Although both photosensitizers proved to have similar properties such as increasing production of reactive oxygen species with the higher concentration, predominance of necrotic mode of death or genotoxicity, the more effective photosensitizer was rose bengal because its EC50 was over 20 times lower for both cell lines than in case of erythrosine B.
- MeSH
- červeň bengálská terapeutické užití MeSH
- erythrosin MeSH
- fibroblasty asociované s nádorem účinky léků účinky záření MeSH
- fotochemoterapie * metody MeSH
- lidé MeSH
- melanom terapie MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- nádory terapie MeSH
- reaktivní formy kyslíku MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Photodynamic therapy (PDT) uses photosensitive substance to provoke a cytotoxic reaction causing a cell damage or cell death. The substances, photosensitizers, are usually derivates of porphyrine or phtalocyanine. Photosensitizers must be activated by light in order to produce reactive oxygen species, mainly singlet oxygen. Sonodynamic therapy (SDT) utilizes ultrasound to enhance a cytotoxic effects of compounds called sonosensitizers. In this study we investigated photodynamic and sonodynamic effect of chloraluminium phtalocyanine disulfonate (ClAlPcS(2)) on HeLa cells. DNA damage, cell viability and reactive oxygen species (ROS) production were assessed to find whether the combination of PDT and SDT inflicts HeLa cells more than PDT alone. We found that the combined therapy increases DNA fragmentation, enhances ROS production and decreases cell survival. Our results indicate that ClAlPcS(2) can act as a sonosentitiser and combined with PDT causes more irreversible changes to the cells resulting in cell death than PDT alone.
- MeSH
- fotochemoterapie * MeSH
- HeLa buňky MeSH
- indoly * MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- organokovové sloučeniny * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Photodynamic therapy is a very promising and constantly evolving diagnostic and therapeutic method that is used mainly for malignant and non-malignant tumors treatment. This study deals with the utilization of zinc photosensitizer (λmax ~ 660 nm) from the group of phthalocyanines in photodynamic therapy. The aim of this study is to evaluate in vitro effect of the 5 Jcm-2 zinc phthalocyanine photosensitizer-mediated photodynamic therapy in EC50 concentration (30 nM) on cytoskeletal apparatus of the tumor cell line-HeLa (cervical cancer cells). For the measurement, the tandem mass spectrometry, atomic force and fluorescent confocal microscopy techniques were used. The results showed, that compared to the control cells zinc-derivative mediated photodynamic therapy caused in HeLa cells significant change of the cell height and extensive cytoskeletal actin rearrangement although the levels of beta actin, gamma actin and F-actin did not change significantly. This is probably caused by decreased level of the ARPC2 actin-related protein which is responsible for actin polymerization. Its level decreased 24 hours after therapy by 56%. The cytoskeletal apparatus is one of the basic cellular structures that provides cell shape, cell division and the intracellular transport. After in vitro 5 Jcm(-2) zinc derivative-mediated photodynamic therapy, the cervical carcinoma cells showed a significant damage of the cytoskeletal structure followed by changes of cell shape leading to cell death. Considering these results and low effective concentration (EC50 = 30 nM), the therapy used is potentially very promising antitumor treatment.
OBJECTIVES: The purpose of this study was to determine toxicity of wastewater from hospitals in the Czech Republic using traditional and alternative toxicological methods. The pilot study comprised weekly dynamics of sewage ecotoxicity of treated wastewater from one hospital in two different seasons. A detailed investigation of wastewater ecotoxicity, genotoxicity and reprotoxicity followed in five different hospitals. METHODS: The seven following bioassays were used in this study: algal growth inhibition test (ISO 8692), Vibrio fischeri test (ISO 11348-2), Daphnia magna acute toxicity test (ISO 6341), Allium cepa assay, Ames test (OECD TG 471), Comet assay and YES/YAS assay. RESULTS: The wastewater ecotoxicity during one week showed no differences in separate working days, however, higher toxicity values were recorded in May compared to November. In the following study, samples from two of the five hospitals were classified as toxic, the others as non toxic. Genotoxicity has not been confirmed in any sample. In several cases, wastewater samples exhibited agonist activity to the estrogen and androgen receptors. CONCLUSION: The study demonstrated different levels of toxicity of treated hospital wastewater. Variable sensitivity of individual bioassays for tested wastewater samples was recognized. A more extensive study including proposal for improvement of hospital wastewater treatment within the Czech Republic can be recommended with the aim to decrease the discharge of toxic chemicals into the local sewage system and the environment.
- MeSH
- Aliivibrio fischeri fyziologie MeSH
- biotest metody MeSH
- česneky fyziologie MeSH
- chemické látky znečišťující vodu analýza toxicita MeSH
- Chlorophyceae fyziologie MeSH
- Daphnia fyziologie MeSH
- nemocnice MeSH
- odpadní voda analýza toxicita MeSH
- odstraňování zdravotnického odpadu MeSH
- pilotní projekty MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
