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
- lidé MeSH
- organokovové sloučeniny * MeSH
- screeningové testy protinádorových léčiv MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- aluminum phthalocyanine disulfonate MeSH Prohlížeč
- indoly * MeSH
- organokovové sloučeniny * MeSH
The aim of the work was early identification of preventable risk factors connected with the consumers usage of products of everyday use, such as cosmetics, toys and children products, and other materials intended for contact with human skin. The risk factor is represented by substances with irritation potential and subsequent possible sensitisation, resulting in negative impact on human physical and psychical health with social and societal consequences. The legislation for cosmetics, chemical substances and other products requires for hazard identification the application of alternative toxicological methods in vitro without the use of animals. For this reason we used a battery of alternative assays in vitro, based on cell cultures. Progressive methods of molecular biology, based on fluorimetry and fluorescence, were employed for identification of early morphological and functional changes on cellular level. Four colorants frequently used in cosmetics (P-WS Caramel, Chlorophyllin, Unicert Red K 7054-J and Unicert Red K 7008-J) were tested on cell line NIH3T3 (mouse fibroblast cell) and 3T3 Balb/c with/without UV irradiation (dose 5 J cm(-2)). Fluorescence methods for the study of cell damage using fluorescence probes offer results for the evaluation of cytotoxicity and cell viability of adherent cells. We detected intracellular production of ROS investigated by molecular probe CM-H(2)DCFDA, which is primarily sensitive to the increased production of hydrogen peroxide or its downstream products. Toxic effects on the cellular level were identified by viability tests using Neutral Red uptake and MTT assay, where the live cells reduce yellow soluble 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to insoluble formazan crystals. The reaction was investigated on mitochondrial membrane of living cells and the type of cell death was determined using Apoptosis detection kit. Cytotoxicity tests revealed health risks of using Chlorophyllin and Unicert Red K 7054-J.
- MeSH
- barvicí látky toxicita MeSH
- buněčná smrt účinky léků MeSH
- buňky BALB 3T3 MeSH
- buňky NIH 3T3 MeSH
- fluorescence MeSH
- fluorometrie metody MeSH
- fototoxická dermatitida etiologie MeSH
- kosmetické přípravky chemie toxicita MeSH
- lidé MeSH
- mitochondriální membrány účinky léků metabolismus MeSH
- myši MeSH
- peroxid vodíku metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- testy toxicity metody MeSH
- ultrafialové záření MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- barvicí látky MeSH
- kosmetické přípravky MeSH
- peroxid vodíku MeSH
- reaktivní formy kyslíku MeSH
Photodynamic therapy (PDT) is an alternative method of tumour treatment. It is based on a photochemical reaction of a photosensitizer, irradiation, and O(2) which converts to cytotoxic (1)O(2) and other forms of reactive oxygen species (ROS). The comet assay (also called single-cell gel electrophoresis, SCGE) is a sensitive, simple and quantitative technique for detection of DNA damage. In our study we investigated the phototoxicity of the two porphyrin photosensitizers, TPPS4 and MgTPPS4, on HeLa cells. Three different radiation doses and six different concentrations of the photosensitizers were used. Our results show that the DNA of the cells treated with the TPPS(4) and MgTPPS(4) at the concentrations higher than 5 μM was highly fragmented indicating a strong phototoxic effect resulting in a cell apoptosis. On the base of our results we can hypothesize that even the irradiation dose of 1 J cm(-2) is sufficient enough to provoke the DNA fragmentation.
- MeSH
- apoptóza účinky léků MeSH
- dávka záření MeSH
- fotochemoterapie metody MeSH
- fotosenzibilizující látky aplikace a dávkování farmakologie MeSH
- fragmentace DNA účinky léků MeSH
- HeLa buňky MeSH
- hořčík chemie MeSH
- kometový test MeSH
- lidé MeSH
- metaloporfyriny aplikace a dávkování chemie farmakologie MeSH
- porfyriny aplikace a dávkování chemie farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fotosenzibilizující látky MeSH
- hořčík MeSH
- magnesium 5,10,15,20-tetrakis(4-sulphophenyl)-porphine dodecahydrate MeSH Prohlížeč
- metaloporfyriny MeSH
- porfyriny MeSH
- tetraphenylporphine sulfonate MeSH Prohlížeč
A measuring system evaluating a Point Spread Function generated in an ultrasonographic image by scanning a spherical target was developed. The target is moved in measuring bath filled by water over scanned volume via 3D computer controlled positioning system. A video signal obtained is converted to digital form and analyzed by original software to derive various objective parameters of the imager as follows: Focal areas in both the azimuth and the elevation directions, Ultrasound scanning lines visualisation, Manufacturer preloaded TGC, Width of the scanning plane, Side lobe levels and Amplification uniformity in the azimuth direction. The method was verified by testing 18 different equipments in 282 measurements. Samples of particular measurement results in form of graphical outputs are included. Medical and physiological impacts of this approach are discussed.
- MeSH
- fantomy radiodiagnostické MeSH
- interpretace obrazu počítačem * MeSH
- počítačové zpracování signálu * MeSH
- reprodukovatelnost výsledků MeSH
- software MeSH
- ultrasonografie přístrojové vybavení metody MeSH
- vylepšení obrazu * MeSH
- zobrazování trojrozměrné * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
