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1.
Plant Dis ; 2023 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-37467129

RESUMO

Viticulture is a traditional branch of agriculture in the Czech Republic. Grapevines (Vitis vinifera L.) are cultivated on more than 18,000 hectares in the wine-growing regions of Bohemia and South Moravia. South Moravia alone accounts for more than 90 % of the total wine-growing area in the country. Grapevine yellows are a complex of diseases associated with the phytoplasma presence. Phytoplasmas of at least five different groups can cause similar symptoms in grapevines, and they can be distinguished only on a molecular basis (EPPO 2016). One of them, the grapevine Flavescence dorée phytoplasma (GFDP), which belongs to the 16SrV group, is listed in Annex II, Part B, of the Commission Implementing Regulation (EU) 2019/2072 of 28 November 2019 as a Union quarantine pest known to occur in the Union territory. Official surveys for GFDP in the Czech Republic have been carried out since 2007. In 2016, the first occurrence of Scaphoideus titanus Ball, 1932, the main vector of GFDP, was reported in the South Moravian Region (EPPO Reporting Service 2016). This is a matter of concern because it indicates that there is a risk of disease dissemination to other geographical locations. In September 2021, a total of 250 samples of V. vinifera (preferentially focused on symptomatic plants) and four samples of the wild plant host Clematis vitalba L. were collected from 50 vineyards in South Moravia. Total DNA was extracted using High Pure PCR Template Preparation Kit (Roche, Basel, Switzerland). For phytoplasma screening, a real-time PCR test for generic detection of phytoplasmas was used (Christensen et al. 2004). Samples evaluated as positive were further tested by PCR using phytoplasma universal P1 and P7 primers (Deng and Hiruki 1991; Schneider et al. 1995), followed by nested PCR using the 16SrV group-specific primers fB1 and rULWS1 (Smart et al. 1996). For identification of 16SrV phytoplasma, sequence analysis of the secY-map genetic locus was performed. Two sets of primers were used: FD9f5/MAPr1 primers for the first PCR and FD9f6/MAPr2 for the nested PCR (Arnaud et al. 2007) with PCRBIO TaqMix (PCR Biosystems, London, UK). The nested PCR products were purified and sequenced (Eurofins Genomics, Ebersberg, Germany). The sequences were compared with sequences from the GenBank database. Phytoplasma of the 16SrV group was detected in three samples: V. vinifera cv. Gewürztraminer with symptoms of leaf reddening with no rolling and no other typical symptoms; C. vitalba L. with leaf curling (Fig. 1A); symptomless C. vitalba. The obtained sequences of the secY-map locus of all three 16SrV-positive samples were identical to the sequence of GFDP, isolate Vv-SI257 (Acc. No. FN811141), detected in grapevine in Tuscany (Italy), which belongs to 16SrV group. The sequence of the V. vinifera cv. Gewürztraminer isolate was submitted to GenBank under Acc. No. OQ185203. This isolate belongs to the Map-FD3 cluster (Fig. 1B), and the genotype identified is M51 (corresponding to FD-C), which has already been found in C. vitalba and outbreaks of Flavescence dorée in grapevines in some other European countries (Malembic-Maher et al. 2020). Based on the abovementioned results, this is the first report of the GFDP in the Czech Republic.

2.
Med Res Rev ; 43(4): 717-774, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-36757198

RESUMO

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.


Assuntos
Neoplasias , Fotoquimioterapia , Humanos , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Fotoquimioterapia/métodos , Neoplasias/tratamento farmacológico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Morte Celular , Espécies Reativas de Oxigênio/metabolismo
3.
Int J Mol Sci ; 23(3)2022 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-35162996

RESUMO

Fluorescent carbon dots (CDs) are potential tools for the labeling of cells with many advantages such as photostability, multicolor emission, small size, rapid uptake, biocompatibility, and easy preparation. Affinity towards organelles can be influenced by the surface properties of CDs which affect the interaction with the cell and cytoplasmic distribution. Organelle targeting by carbon dots is promising for anticancer treatment; thus, intracellular trafficking and cytotoxicity of cationic CDs was investigated. Based on our previous study, we used quaternized carbon dots (QCDs) for treatment and monitoring the behavior of two human cancer cell MCF-7 and HeLa lines. We found similarities between human cancer cells and mouse fibroblasts in the case of QCDs uptake. Time lapse microscopy of QCDs-labeled MCF-7 cells showed that cells are dying during the first two hours, faster at lower doses than at higher ones. QCDs at a concentration of 100 µg/mL entered into the nucleus before cellular death; however, at a dose of 200 µg/mL, blebbing of the cellular membrane occurred, with a subsequent penetration of QCDs into the nuclear area. In the case of HeLa cells, the dose-depended effect did not happen; however, the labeled cells were also dying in mitosis and genotoxicity occurred nearly at all doses. Moreover, contrasted intracellular compartments, probably mitochondria, were obvious after 24 h incubation with 100 µg/mL of QCDs. The levels of reactive oxygen species (ROS) slightly increased after 24 h, depending on the concentration, thus the genotoxicity was likely evoked by the nanomaterial. A decrease in viability did not reach IC 50 as the DNA damage was probably partly repaired in the prolonged G0/G1 phase of the cell cycle. Thus, the defects in the G2/M phase may have allowed a damaged cell to enter mitosis and undergo apoptosis. The anticancer effect in both cell lines was manifested mainly through genotoxicity.


Assuntos
Carbono/farmacocinética , Fibroblastos/citologia , Neoplasias/metabolismo , Pontos Quânticos/química , Espécies Reativas de Oxigênio/metabolismo , Imagem com Lapso de Tempo/métodos , Animais , Transporte Biológico , Carbono/química , Carbono/farmacologia , Linhagem Celular , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Células HeLa , Humanos , Células MCF-7 , Camundongos , Neoplasias/tratamento farmacológico , Neoplasias/genética , Imagem Óptica
4.
Int J Mol Sci ; 22(11)2021 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-34070594

RESUMO

It is important to understand the nanomaterials intracellular trafficking and distribution and investigate their targeting into the nuclear area in the living cells. In our previous study, we firstly observed penetration of nonmodified positively charged carbon dots decorated with quaternary ammonium groups (QCDs) into the nucleus of mouse NIH/3T3 fibroblasts. Thus, in this work, we focused on deeper study of QCDs distribution inside two healthy mouse NIH/3T3 and L929 cell lines by fluorescence microspectroscopy and performed a comprehensive cytotoxic and DNA damage measurements. Real-time penetration of QCDs across the plasma cell membrane was recorded, concentration dependent uptake was determined and endocytic pathways were characterized. We found out that the QCDs concentration of 200 µg/mL is close to saturation and subsequently, NIH/3T3 had a different cell cycle profile, however, no significant changes in viability (not even in the case with QCDs in the nuclei) and DNA damage. In the case of L929, the presence of QCDs in the nucleus evoked a cellular death. Intranuclear environment of NIH/3T3 cells affected fluorescent properties of QCDs and evoked fluorescence blue shifts. Studying the intracellular interactions with CDs is essential for development of future applications such as DNA sensing, because CDs as DNA probes have not yet been developed.


Assuntos
Carbono , Ciclo Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Núcleo Celular/metabolismo , Fibroblastos/metabolismo , Pontos Quânticos , Animais , Carbono/química , Carbono/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Camundongos , Microscopia de Fluorescência , Células NIH 3T3 , Pontos Quânticos/química , Pontos Quânticos/uso terapêutico
5.
Int J Pharm ; 528(1-2): 280-286, 2017 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-28602801

RESUMO

Silver nanoparticles (AgNPs) have been widely studied for their beneficial antimicrobial effect and have been considered by some to be a safe ingredient, as penetration of metal nanoparticles through the skin in vivo has not been proven. However, AgNPs are becoming a commonly applied nanomaterial for surface modifications of medical products which come into contact with damaged skin. In our experiments, we tested two commercially available AgNPs samples manufactured by electrolysis. AFM was used to characterize tested AgNPs morphology and their mean particle size which was assessed as 30.6nm and 20.4nm. An important mechanism of AgNPs cytotoxicity is generation of reactive oxygen species (ROS), chemically reactive species containing oxygen. Although ROS occur in cell metabolism naturally, their overproduction can induce oxidative stress - imbalance between production and antioxidant defenses. This can be associated with cytotoxicity and DNA damage. Conventional in vitro tests were used to evaluate the cytotoxic potential and DNA damage in BJ human fibroblasts cell lines. We found that both tested AgNPs samples induced ROS generation and caused the DNA damage in fibroblasts. One of the key concerns about the association with cytotoxic or genotoxic responses of nanoparticles is the capability of these materials to penetrate through cellular membrane. Cellular uptake studies were performed using Raman imaging as a label-free microscopic technique. In combination with a univariate image analysis, results demonstrate cellular uptake and distribution of the AgNPs which were taken up by BJ cells within 24h of incubation in a growth medium. The study demonstrates the potential of Raman imaging to unambiguously identify and localize AgNPs in fixed cells.


Assuntos
Fibroblastos/metabolismo , Nanopartículas Metálicas , Prata/metabolismo , Linhagem Celular , Humanos , Espécies Reativas de Oxigênio/metabolismo , Análise Espectral Raman
6.
Cent Eur J Public Health ; 24(3): 234-240, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27755863

RESUMO

AIM: Natural or artificial substances have become an inseparable part of our lives. It is questionable whether adequate testing has been performed in order to ensure these substances do not pose a serious health risk. The principal aim of our research was to clarify the potential risk of adding essential oils to food, beverages and cosmetic products. METHODS: The toxicity of substances frequently employed in cosmetics, aromatherapy and food industry (bergamot oil, Litsea cubeba oil, orange oil, citral) were investigated using cell line NIH3T3 (mouse fibroblasts) with/without UV irradiation. The MTT assay was used to estimate the cell viability. Reactive oxygen species (ROS) which are products of a number of natural cellular processes such as oxygen metabolism and inflammation were measured to determine the extent of cellular stress. DNA damage caused by strand breaks was examined by comet assay. RESULTS: MTT test determined EC50 values for all tested substances, varying from 0.0023% v/v for bergamot oil to 0.018% v/v for citral. ROS production measurement showed that UV radiation induces oxidative stress to the cell resulting in higher ROS production compared to the control and non-irradiated samples. Comet assay revealed that both groups (UV, without UV) exert irreversible DNA damage resulting in a cell death. CONCLUSIONS: Our findings suggest that even low concentrations (lower than 0.0464% v/v) of orange oil can be considered as phototoxic (PIF value 8.2) and probably phototoxic for bergamot oil (PIF value 4.6). We also found significant changes in the cell viability, the ROS production and the DNA after the cells were exposed to the tested chemicals. Even though these substances are widely used as antioxidants it should be noted that they present a risk factor and their use in cosmetic and food products should be minimized.


Assuntos
Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos da radiação , Litsea/toxicidade , Monoterpenos/toxicidade , Células NIH 3T3/efeitos dos fármacos , Células NIH 3T3/efeitos da radiação , Óleos de Plantas/toxicidade , Raios Ultravioleta , Monoterpenos Acíclicos , Animais , Ensaio Cometa , Dano ao DNA , Dermatite Fototóxica , Camundongos , Estresse Oxidativo , Espécies Reativas de Oxigênio
7.
Food Chem Toxicol ; 96: 50-61, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27456126

RESUMO

Silver nanoparticles (AgNPs) are the most frequently applied nanomaterials. In our experiments, we tested AgNPs (size 27 nm) manufactured by the Tollens process. Physico-chemical methods (TEM, DLS, AFM and spectrophotometry) were used for characterization and imaging of AgNPs. The effects of AgNPs and Ag(+) were studied in two experimental models (plant and mammalian cells). Human keratinocytes (SVK14) and mouse fibroblasts (NIH3T3) cell lines were selected to evaluate the cytotoxicity and genotoxicity effect on mammalian cells. Higher sensitivity to AgNPs and Ag(+) was observed in NIH3T3 than in SVK14 cells. AgNPs accumulated in the nucleus of NIH3T3 cells, caused DNA damage and increased the number of apoptotic and necrotic cells. Three genotypes of Solanum spp. (S. lycopersicum cv. Amateur, S. chmielewskii, S. habrochaites) were selected to test the toxicity of AgNPs and Ag(+) on the plant cells. The highest values of peroxidase activity and lipid peroxidation were recorded after the treatment of S. habrochaites genotype with AgNPs. Increased ROS levels were likely the reason for observed damaged membranes in S. habrochaites. We found that the cytotoxic and genotoxic effects of AgNPs depend not only on the characteristics of nanoparticles, but also on the type of cells that are treated with AgNPs.


Assuntos
Apoptose/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Prata/farmacologia , Solanum lycopersicum/citologia , Animais , Western Blotting , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Ensaio Cometa , Humanos , Técnicas In Vitro , Solanum lycopersicum/efeitos dos fármacos , Nanopartículas Metálicas/química , Camundongos , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Prata/química , Análise Espectral Raman
8.
Cell Biochem Biophys ; 74(3): 419-25, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27324041

RESUMO

Metalloporphyrins are an important group of sensitizers with a porphyrin skeleton. Their photophysical properties are significantly affected by the nature of the central ion. In this work, we focus on the mechanical properties of a cervix carcinoma cell line which underwent photodynamic treatment (PDT) with MgTPPS4 photosensitzer. Atomic force microscopy alongside confocal microscopy was used to quantify and qualify the structural characteristics before and after PDT. Cells before PDT showed a fine actin network and higher elasticity with the median of Young modulus 12.2 kPa. After PDT, the median of Young modulus was 13.4 kPa and a large redistribution in the actin network was observed.


Assuntos
Citoesqueleto/efeitos dos fármacos , Metaloporfirinas/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Citoesqueleto/fisiologia , Citoesqueleto/efeitos da radiação , Módulo de Elasticidade , Células HeLa , Humanos , Luz , Metaloporfirinas/química , Microscopia de Força Atômica , Microscopia Confocal , Fármacos Fotossensibilizantes/química
9.
Carbohydr Polym ; 144: 68-75, 2016 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-27083794

RESUMO

Hyaluronan (HA) films exhibit properties suitable for various biomedical applications, but the solubility of HA limits their use in aqueous environments. Therefore, we developed water insoluble films based on palmitoyl esters of HA (pHA). Films were prepared from pHA samples with various degrees of substitution (DS) and molecular weights and their mechanical properties and swelling were characterized. Additionally, scanning electron microscopy and atomic force microscopy were used for visualization. Despite being prepared by solution casting, the films had a very smooth surface and were homogeneous in thickness. The film properties were in accordance with the polymer DS and molecular weight, enabling to tailor them for future applications by choosing a suitable pHA material. The behavior of the films toward cells was assessed in vitro. All films were non-cytotoxic and showed no adhesion of cells. These results show that the developed films are suitable candidates for various biomedical applications such as tissue engineering or wound healing.


Assuntos
Ácido Hialurônico/química , Palmitatos/química , Acilação , Animais , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Citometria de Fluxo , Humanos , Ácido Hialurônico/análogos & derivados , Ácido Hialurônico/toxicidade , Camundongos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Palmitatos/toxicidade , Solubilidade , Células Swiss 3T3 , Resistência à Tração , Água/química
10.
Food Chem Toxicol ; 85: 20-30, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26518667

RESUMO

Commercially manufactured nanomaterials are used massively for modification of products of everyday use, including products intended for children. Therefore their potential risks have to be ultimately studied. Aside from toxicity of nanomaterials with known specific parameters, the end-consumer is potentially endangered by materials with unknown specification. Commercially available products are not usually accompanied by parameter/specification sheet providing the consumer with sufficient chemico-physical parameters allowing the evaluation of possible toxic effects. The aim of this work was to evaluate the declared parameters of commercially available TiO2 and Ag NPs employing chemico-physical methods and consequently in vitro cytotoxicity and genotoxicity tests performed on non-cancer cell lines. Based on the results of our complex study we can conclude that the data provided by the producers are not in good agreement with the performed measurements. Furthermore, all tested NPs penetrated into the SVK14 cells and all NPs had significant effect on the kinetics of ROS production in all cell lines (note: the ROS production has not been established as the major mechanism of cell damage elicited by Ag NPs). The study revealed greater cytotoxic potential of Ag NPs in comparison with TiO2 NPs and all of the studied NPs caused significant DNA damage.

11.
Int J Pharm ; 496(2): 878-85, 2015 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-26456248

RESUMO

Silver nanoparticles (AgNPs) have been used for decades as anti-bacterial agents in various industrial fields such as cosmetics, health industry, food storage, textile coatings and environmental applications, although their toxicity is not fully recognized yet. Antimicrobial and catalytic activity of AgNPs depends on their size as well as structure, shape, size distribution, and physico-chemical environment. The unique properties of AgNPs require novel or modified toxicological methods for evaluation of their toxic potential combined with robust analytical methods for characterization of nanoparticles applied in relevant vehicles, e.g., culture medium with/without serum and phosphate buffered saline.


Assuntos
Nanopartículas Metálicas/química , Prata/química , Difusão Dinâmica da Luz , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Veículos Farmacêuticos
12.
Food Chem Toxicol ; 82: 106-15, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25846500

RESUMO

Commercially manufactured nanomaterials are used massively for modification of products of everyday use, including products intended for children. Therefore their potential risks have to be ultimately studied. Aside from toxicity of nanomaterials with known specific parameters, the end-consumer is potentially endangered by materials with unknown specification. Commercially available products are not usually accompanied by parameter/specification sheet providing the consumer with sufficient chemico-physical parameters allowing the evaluation of possible toxic effects. The aim of this work was to evaluate the declared parameters of commercially available TiO2 and Ag NPs employing chemico-physical methods and consequently in vitro cytotoxicity and genotoxicity tests performed on non-cancer cell lines. Based on the results of our complex study we can conclude that the data provided by the producers are not in good agreement with the performed measurements. Furthermore, all tested NPs penetrated into the SVK14 cells and all NPs had significant effect on the kinetics of ROS production in all cell lines (note: the ROS production has not been established as the major mechanism of cell damage elicited by Ag NPs). The study revealed greater cytotoxic potential of Ag NPs in comparison with TiO2 NPs and all of the studied NPs caused significant DNA damage.


Assuntos
Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Titânio/toxicidade , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular/efeitos dos fármacos , Linhagem Celular/metabolismo , Ensaio Cometa , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanopartículas Metálicas/química , Camundongos , Microscopia de Força Atômica , Células NIH 3T3/efeitos dos fármacos , Tamanho da Partícula , Espécies Reativas de Oxigênio/metabolismo , Prata/química , Prata/farmacocinética , Espectrofotometria Atômica , Análise Espectral Raman , Titânio/farmacocinética
13.
Int J Nanomedicine ; 10: 949-61, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25673990

RESUMO

One of the promising strategies for improvement of cancer treatment is based on magnetic drug delivery systems, thus avoiding side effects of standard chemotherapies. Superparamagnetic iron oxide (SPIO) nanoparticles have ideal properties to become a targeted magnetic drug delivery contrast probes, named theranostics. We worked with SPIO condensed colloidal nanocrystal clusters (MagAlg) prepared through a new soft biomineralization route in the presence of alginate as the polymeric shell and loaded with doxorubicin (DOX). The aim of this work was to study the in vitro cytotoxicity of these new MagAlg-DOX systems on mouse fibroblast and breast carcinoma cell lines. For proper analysis and understanding of cell behavior after administration of MagAlg-DOX compared with free DOX, a complex set of in vitro tests, including production of reactive oxygen species, comet assay, cell cycle determination, gene expression, and cellular uptake, were utilized. It was found that the cytotoxic effect of MagAlg-DOX system is delayed compared to free DOX in both cell lines. This was attributed to the different mechanism of internalization of DOX and MagAlg-DOX into the cells, together with the fact that the drug is strongly bound on the drug nanocarriers. We discovered that nanoparticles can attenuate or even inhibit the effect of DOX, particularly in the tumor MCF7 cell line. This is a first comprehensive study on the cytotoxic effect of DOX-loaded SPIO compared with free DOX on healthy and cancer cell lines, as well as on the induced changes in gene expression.


Assuntos
Antineoplásicos , Sobrevivência Celular/efeitos dos fármacos , Coloides , Doxorrubicina , Nanopartículas de Magnetita , Animais , Antineoplásicos/química , Antineoplásicos/toxicidade , Coloides/química , Coloides/toxicidade , Doxorrubicina/química , Doxorrubicina/toxicidade , Humanos , Células MCF-7 , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/toxicidade , Camundongos , Células NIH 3T3
14.
Indian J Biochem Biophys ; 51(1): 19-28, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24791413

RESUMO

Photodynamic treatment (PDT) in combination with sonodynamic treatment (SDT) can be used as suitable methods to treat malignant and benign diseases or combat resistant bacteria. Both methods affect the production of reactive oxygen species (ROS). On the other hand, antioxidants are useful for cell protection against ROS. This work was aimed to study the effect of PDT and SDT treatments on the HeLa cell line using antioxidant Pronalen Sensitive Skin as a protection from free radicals in the cells. We evaluated the effect of sensitizer ClAlPcS2 using battery of in vitro methods, including MTT assay, kinetic production of ROS, mitochondrial membrane potential change, type of cell death and microscopic analysis. Ultrasound treatment was observed to increase the production of ROS, only in combination with PDT, particularly at higher concentrations of ClAlPcS2. The added antioxidant acts as protection against free radicals and has potential as a dietary supplement against aging or free radicals. The results of study suggested that ClAlPcS2 could be used as a potential photosensitizer for treatment of a specific type of cancers.


Assuntos
Antioxidantes/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Terapia por Ultrassom , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Células HeLa , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos da radiação , Necrose , Espécies Reativas de Oxigênio/metabolismo
15.
Artigo em Inglês | MEDLINE | ID: mdl-23073531

RESUMO

BACKGROUND: Bacterial resistance to antibiotics is a constantly growing challenge. Photodynamic therapy (PDT) offers a new approach to the treatment of bacterial and viral diseases. The aim of this study was to compare the efficacy of photosensitizers used in PDT applied to cell lines and bacterial strains. METHODS: We tested the cytotoxicity and phototoxicity of 3 photosensitizers: TPPS4, ZnTPPS4 and TMPyP applied to the NIH3T3 cell line using two established methods for measuring ROS production and, MTT viability assay. Bacterial viability was determined spectrophotometrically over 24 h following PDT. RESULTS: The most efficient photosensitiser was TMPyP as it reduced the viability of the NIH3T3 cell line by more than 85%. In general, the photosensitisers were more phototoxic to the two Gram-positive bacterial strains, Enterococcus faecalis and Staphylococcus aureus. The viability of E. faecalis was reduced to 78 % by a dose radiation 0.5 J/cm(2) and concentration of TMPyP 1.562 µmol/L. The viability of bacterium S. aureus was reduced to 23 % when exposed to a radiation dose 0.5 J/cm(2) and 100 µmol/L concentration of ZnTPPS4. The highest viability decrease (15 %) for Pseudomonas aeruginosa was caused by 0.5 J/cm(2) radiation dose and 50 µmol/L TMPyP concentration. Escherichia coli proved to be PDT resistant as the bacterial viability was higher than 90%. CONCLUSIONS: The goal of the present study was to test the efficiency of photosensitizers on the NIH 3T3 cell line and bacterial cells. Subsequently we would like to study effectiveness of photosensitizers bound to carriers (for example cyclodextrins) on other cell line and bacterial strain.


Assuntos
Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/efeitos da radiação , Células NIH 3T3/efeitos dos fármacos , Células NIH 3T3/efeitos da radiação , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/efeitos da radiação , Escherichia coli/efeitos dos fármacos , Escherichia coli/efeitos da radiação , Metaloporfirinas/farmacologia , Camundongos , Porfirinas/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos da radiação , Espécies Reativas de Oxigênio/análise , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/efeitos da radiação
16.
Microbiol Res ; 169(2-3): 163-70, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-23899404

RESUMO

Photodynamic therapy is usually used against malignant and non-malignant tumors. Nowadays, due to resistance of bacterial strains, we are looking for a new antimicrobial strategy to destroy bacteria with minimal invasive consequences. The worldwide increase in antibiotic resistance among different classes of gram-positive and gram-negative bacteria has led to the search for alternative anti-microbial therapies such as antimicrobial PDT (aPDT). Development antimicrobial technology combines a nontoxic compound, called photosensitizer, visible light of the appropriate wavelength, and the generation of reactive oxygen species. In this work, the photosensitizers TMPyP and ZnTPPS4 are investigated for photodynamic and antimicrobial photodynamic therapy. We tested these two porphyrins on two cell lines and two bacterial strains to compare effectiveness. In addition, we applied photosensitizers bound in the complex created with hp-ß-cyclodextrin. The light-emitting diodes were used at the doses 0, 1, 5, 10 J/cm(2) for cells and 0, 150 J/cm(2) for bacteria. Tested concentrations for cells and microbes were from 0.5 to 50 µM and from 0.78 to 100 µM, respectively. From this work it can be concluded that TMPyP is a promising compound both in aPDT and in PDT, particularly in contrast to ZnTPPS4, which was efficient only in PDT. Furthermore, the eradication of gram-positive bacteria is possible only with higher concentrations of ZnTPPS4.


Assuntos
Ciclodextrinas/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/efeitos da radiação , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/efeitos da radiação , Linhagem Celular , Ciclodextrinas/química , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/microbiologia , Humanos , Luz , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Porfirinas/química , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia
17.
Gen Physiol Biophys ; 32(3): 337-46, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23817636

RESUMO

High resolution imaging of biological structures and changes induced by various agents such as drugs and toxins is commonly performed by fluorescence and electron microscopy (EM). Although high-resolution imaging is possible with EM, the requirements for fixation and staining of samples for image contrast severely limits the study of living organisms. Atomic force microscopy (AFM), on the other hand, is capable of simultaneous nanometer spatial resolution and piconewton force detection, allowing detailed study of cell surface morphology and monitoring cytomechanical information. We present a method that images and studies mechanically characterized cells using AFM. We used a HeLa cell line (cervix carcinoma cell), which is sensitive to photodynamic treatment (PDT); growth media as a scanning surrounding; atomic force microscopy NT-MDT Aura for cytomechanical measurement; and scanning electron microscope Hitachi Su 6600 for control images of the cells. The modulus of elasticity for intact and photodynamically damaged cells can indicate mechanical changes to the main properties of cells. Cell elasticity changes can provide information on the degree or value of cell damage, for example after PDT. Measurements were carried out on approximately sixty cells, including three independent experiments on a control group and on sixty cells in a photodamaged group. Cells before PDT show higher elasticity: the median of Young´s modulus on the nucleus was 35.283 kPa and outside of the nucleus 107.442 kPa. After PDT, the median of Young's modulus on the nucleus was 61.144 kPa and outside of the nucleus was 193.605 kPa.


Assuntos
Fotoquimioterapia , Fenômenos Biomecânicos , Módulo de Elasticidade/efeitos dos fármacos , Módulo de Elasticidade/efeitos da radiação , Células HeLa , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura
18.
J Biomed Mater Res A ; 101(10): 2915-24, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23529774

RESUMO

The modification of implant surface situated in the area of peri-implant sulcus has important role in bacterial and cell adhesion. Six different chemically and physically modified titanium discs were prepared: glazed (Tis-MALP), unglazed (Tis-O), unglazed and alkali-etched (Tis-OA), unglazed and coated with ZrN (Tis-OZ), unglazed, sand blasted, and acid etched (Tis-OPAE), and unglazed, sand blasted, acid, and alkali etched (Tis-OPAAE). Analysis of surface topography was determined using scanning electron microscopy and atomic force microscopy (AFM). Biocompatibility of gingival fibroblasts was characterized by the production of tumor necrosis factor alpha, collagen I, matrix metalloproteinase 2 (MMP-2) after 24 and 72 h and expression of α3 ß1 integrin and vinculin using enzyme-linked immunosorbent assay (ELISA) or modified ELISA after 6 and 24 h. Microorganism adhesion (five bacterial strains) and biofilm formation was also evaluated. The adhesion of bacteria and gingival fibroblasts was significantly higher on titanium disc Tis-OPAAE and biofilm formation on the same surface for Streptococcus mutans, Streptococcus gordonii, and Streptococcus intermedius. The gingival fibroblasts on Tis-OPAAE disc had also significantly lower production of MMP-2. The collagen production was significantly lower on all surfaces with roughness higher than 0.2 µm. This study confirmed that the titanium disc with the surface roughness 3.39 µm (Tis-OPAAE) supported the adhesion of bacterial strains as well as gingival fibroblasts.


Assuntos
Materiais Biocompatíveis/farmacologia , Fibroblastos/citologia , Fibroblastos/microbiologia , Gengiva/citologia , Teste de Materiais , Streptococcus/citologia , Titânio/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Adesão Celular/efeitos dos fármacos , Colágeno Tipo I/biossíntese , Implantes Dentários/microbiologia , Fibroblastos/efeitos dos fármacos , Adesões Focais/efeitos dos fármacos , Adesões Focais/metabolismo , Humanos , Integrina alfa3beta1/metabolismo , Metaloproteinase 2 da Matriz/biossíntese , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Espectrometria por Raios X , Streptococcus/efeitos dos fármacos , Streptococcus/fisiologia , Fator de Necrose Tumoral alfa/biossíntese , Vinculina/metabolismo
19.
Chem Commun (Camb) ; 48(93): 11398-400, 2012 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-23066527

RESUMO

Core-shell hydrophilic superparamagnetic iron oxide (SPIO) nanoparticles, surface functionalized with either terephthalic acid or 2-amino terephthalic acid, showed large negative MRI contrast ability, validating the advantage of using low molecular weight and π-conjugated canopies for engineering functional nanostructures with superior performances.


Assuntos
Meios de Contraste/química , Desenho de Fármacos , Compostos Férricos/química , Imageamento por Ressonância Magnética/métodos , Imãs/química , Animais , Meios de Contraste/toxicidade , Compostos Férricos/toxicidade , Imãs/toxicidade , Camundongos , Células NIH 3T3 , Propriedades de Superfície
20.
Artigo em Inglês | MEDLINE | ID: mdl-22837129

RESUMO

BACKGROUND: Photodynamic therapy (PDT) is a new modality in cancer treatment. It is based on the tumour-selective accumulation of a photosensitizer followed by irradiation with light of a specific wavelength. PDT is becoming widely accepted owing to its relative specificity and selectivity along with absence of the harmful side-effects of chemo and radiotherapy. There are three known distinct mechanisms of tumour destruction following PDT, generation of reactive oxygen species which can directly kill tumour cells, tumour vascular shutdown which can independently lead to tumour destruction via lack of oxygen and nutrients and thirdly enhanced antitumour immunity. METHODS: A review based on the literature acquired from the PubMed database from 1983 with a focus on the enhanced antitumour immunity effects of PTD. RESULTS AND CONCLUSION: Tumour cell death is accompanied by the release of a large number of inflammatory mediators. These induce a non-specific inflammatory response followed by gradual adaptive antitumour immunity. Further, a combination of PDT with the immunological approach has the potential to improve PDT efficiency and increase the cure rate. This short review covers specific methods for achieving these goals.


Assuntos
Neoplasias/tratamento farmacológico , Fotoquimioterapia , Imunidade Adaptativa/efeitos dos fármacos , Animais , Ativação do Complemento , Citocinas/metabolismo , Humanos , Tolerância Imunológica , Imunidade Inata/efeitos dos fármacos , Neoplasias/imunologia , Estresse Oxidativo , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico
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