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.
- MeSH
- cytoskelet účinky léků fyziologie účinky záření MeSH
- fotosenzibilizující látky chemie farmakologie MeSH
- HeLa buňky MeSH
- konfokální mikroskopie MeSH
- lidé MeSH
- metaloporfyriny chemie farmakologie MeSH
- mikroskopie atomárních sil MeSH
- modul pružnosti MeSH
- světlo MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
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 principalAIM 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.
- MeSH
- buňky NIH 3T3 účinky léků účinky záření MeSH
- fibroblasty účinky léků účinky záření MeSH
- fototoxická dermatitida MeSH
- kometový test MeSH
- Litsea toxicita MeSH
- monoterpeny toxicita MeSH
- myši MeSH
- oleje rostlin toxicita MeSH
- oxidační stres MeSH
- poškození DNA MeSH
- reaktivní formy kyslíku MeSH
- ultrafialové záření * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Photodynamic therapy (PDT) is based on the tumor-selective accumulation of photosensitizer followed by irradiation with light of an appropriate wavelength. After irradiation and in the presence of oxygen, photosensitizer induces cellular damage. The aim of this study was to evaluate effects of two photosensitizers TMPyP and ClAlPcS2 on cell lines to obtain better insight into their mechanisms of action. We determined cell viability, reactive oxygen species (ROS) generation and changes in expression levels of two important early response genes, C-MYC and C-FOS, on tumor MCF7 (human breast adenocarcinoma) and G361 (human melanoma) cell lines and non-tumor BJ cell line (human fibroblast) after photodynamic reaction with TMPyP and ClAlPcS2 as photosensitizers. In addition TMPyP and ClAlPcS2 cellular uptake and clearance and antioxidant capacity of the mentioned cell lines were investigated. We found appropriate therapeutic doses and confirmed that both tested photosensitizers are photodynamically efficient in treatment used cells in vitro. TMPyP is more efficient; it had higher ROS production and toxicity after irradiation by intermediate therapeutic doses than ClAlPcS2. We revealed that both TMPyP and ClAlPcS2-PDT increased C-FOS expression on tumor cell lines (G361 and MCF7), but not on non-tumor BJ cell line. Conversely, both TMPyP and ClAlPcS2-PDT decreased C-MYC expression on non-tumor BJ cell line but not on tumor cell lines. As first we tested these photosensitizers in such extent and we believe that it can help to better understand mechanisms of PDT and increase its efficiency and applicability.
- MeSH
- antioxidancia metabolismus MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky chemie terapeutické užití toxicita MeSH
- indoly chemie terapeutické užití toxicita MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie MeSH
- organokovové sloučeniny chemie terapeutické užití toxicita MeSH
- porfyriny chemie terapeutické užití toxicita MeSH
- protoonkogenní proteiny c-fos metabolismus MeSH
- protoonkogenní proteiny c-myc metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- světlo MeSH
- upregulace účinky léků účinky záření MeSH
- viabilita buněk účinky léků účinky záření MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
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.
- MeSH
- antitumorózní látky * chemie toxicita MeSH
- buňky NIH 3T3 MeSH
- doxorubicin * chemie toxicita MeSH
- koloidy * chemie toxicita MeSH
- lidé MeSH
- magnetické nanočástice * chemie toxicita MeSH
- MFC-7 buňky MeSH
- myši MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- photodynamická terapie,
- MeSH
- cévy fyziologie růst a vývoj sekrece účinky léků MeSH
- exprese genu účinky léků MeSH
- fibroblastové růstové faktory MeSH
- fotochemoterapie využití MeSH
- fotosenzibilizující látky MeSH
- inhibitory angiogeneze * aplikace a dávkování terapeutické užití MeSH
- kyselina salicylová aplikace a dávkování MeSH
- látky indukující angiogenezi MeSH
- lidé MeSH
- nádory * terapie MeSH
- prospektivní studie MeSH
- TNF-alfa toxicita MeSH
- vaskulární endoteliální růstové faktory sekrece MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- grafy a diagramy MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- hASC+HUVEC co-kultura,
- MeSH
- antiinfekční látky terapeutické užití MeSH
- fotochemoterapie využití MeSH
- fotosenzibilizující látky aplikace a dávkování terapeutické užití MeSH
- inhibitory angiogeneze * fyziologie MeSH
- kyselina salicylová * aplikace a dávkování terapeutické užití MeSH
- lidé MeSH
- nádory terapie MeSH
- prospektivní studie MeSH
- vaskulární endoteliální růstové faktory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- techniky in vitro MeSH
BACKGROUND: Photodynamic therapy (PDT) is linked with oxidative damage of biomolecules causing significant impairment of essential cellular functions that lead to cell death. It is the reason why photodynamic therapy has found application in treatment of different oncological, cardiovascular, skin and eye diseases. Efficacy of PDT depends on combined action of three components; sensitizer, light and oxygen. In the present study, we examined whether higher partial pressure of oxygen increases lethality in HeLa cell lines exposed to light in the presence of chloraluminium phthalocyanine disulfonate (ClAlPcS2). METHODS: ClAlPcS2- sensitized HeLa cells incubated under different oxygen conditions were exposed to PDT. Production of singlet oxygen ((1)O2) and other forms of reactive oxygen species (ROS) as well as changes in mitochondrial membrane potential were determined by appropriately sensitive fluorescence probes. The effect of PDT on HeLa cell viability under different oxygen conditions was quantified using the standard methylthiazol tetrazolium (MTT) test. RESULTS: At the highest oxygen concentration of 28 ± 2 mg/l HeLa cells were significantly more sensitive to light-activated ClAlPcS2 (EC50=0.29 ± 0.05 μM) in comparison to cells incubated at lower oxygen concentrations of 8 ± 0.5 and 0.5 ± 0.1 mg/l, where the half maximal effective concentration was 0.42 ± 0.06 μM and 0.94 ± 0.14 μM, respectively. Moreover, we found that the higher presence of oxygen is accompanied with higher production of singlet oxygen, a higher rate of type II photodynamic reactions, and a significant drop in the mitochondrial membrane potential. CONCLUSION: These results demonstrate that the photodynamic effect in cervical cancer cells utilizing ClAlPcS2 significantly depends on oxygen level.
- MeSH
- fotochemoterapie * MeSH
- fotosenzibilizující látky farmakologie MeSH
- HeLa buňky MeSH
- indoly farmakologie MeSH
- kyslík analýza farmakologie MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- parciální tlak MeSH
- reaktivní formy kyslíku metabolismus MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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-β-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.
- MeSH
- buněčné linie MeSH
- cyklodextriny chemie farmakologie MeSH
- Escherichia coli účinky léků účinky záření MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky chemie farmakologie MeSH
- infekce vyvolané Escherichia coli farmakoterapie mikrobiologie MeSH
- lidé MeSH
- porfyriny chemie farmakologie MeSH
- stafylokokové infekce farmakoterapie mikrobiologie MeSH
- Staphylococcus aureus účinky léků účinky záření MeSH
- světlo MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
