Cobalt oxide nanoparticles were prepared via green chemistry route and fully characterized by Field Emission Scanning Electron Microscope (FESEM), Energy-dispersive X-ray spectroscopy (EDAX), X-ray diffraction (XRD), High-resolution transmission electron microscopy (HRTEM) and Transmission electron microscopy (TEM) analyses; the CoO and Co3O4 nanoparticles, in sheet-shaped cobalt oxide form, ensued simultaneously in one step. The varying concentrations of NPs were analyzed via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test on the cancer cell line (U87) which revealed that with increasing concentration of cobalt oxide nanoparticles, the survival rate of U87 tumor cells decreases; IC50 of nanoparticles being ~ 55 µg/ml-1.
- MeSH
- antibakteriální látky chemie MeSH
- antitumorózní látky farmakologie MeSH
- difrakce rentgenového záření MeSH
- inhibiční koncentrace 50 MeSH
- kobalt chemie MeSH
- koncentrace vodíkových iontů MeSH
- kovové nanočástice chemie MeSH
- lidé MeSH
- magnetismus MeSH
- mikrobiální testy citlivosti MeSH
- nádorové buněčné linie MeSH
- nanomedicína metody MeSH
- nanotechnologie metody MeSH
- oxidy chemie MeSH
- povrchově aktivní látky MeSH
- rostlinné extrakty MeSH
- rozmarýn MeSH
- rozpustnost MeSH
- spektroskopie infračervená s Fourierovou transformací MeSH
- technologie zelené chemie metody MeSH
- teplota MeSH
- tetrazoliové soli chemie MeSH
- thiazoly chemie MeSH
- transmisní elektronová mikroskopie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Skin explants are a suitable model which can replace dermatological experiments on animals or human volunteers. In this study, we searched for a fast, cheap and reproducible method for screening skin explant viability after treatment with UVA radiation or/and chemical agents. We compared frequently used methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), neutral red (NR) and lactate dehydrogenase (LDH) activity assay with a rarely used 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) assay for the evaluation of UVA radiation and/or chlorpromazine and 8-methoxypsoralen effect as model agents. Histological analysis of skin explants was also performed by a simple haematoxylin-eosin method. Only the TTC assay was able to show the toxicity of model agents in a dose- and concentration-dependent manner. LDH assay was partially able to demonstrate results comparable to the TTC method, however, the agents' effect was less pronounced. The MTT and NR assays completely failed in the evaluation. Haematoxylin-eosin staining showed discrete structural changes in samples treated with UVA alone and CPZ+UVA, but only after 48h. Therefore, the method is not useful for screening of toxic or phototoxic effects either. In conclusion, the TTC assay was the most suitable for the evaluation of toxicity or phototoxicity in ex vivo skin.
- MeSH
- biotest * MeSH
- chlorpromazin toxicita MeSH
- kultivované buňky MeSH
- kůže cytologie účinky léků patologie MeSH
- L-laktátdehydrogenasa metabolismus MeSH
- lidé MeSH
- methoxsalen toxicita MeSH
- techniky in vitro MeSH
- tetrazoliové soli chemie MeSH
- ultrafialové záření * MeSH
- viabilita buněk účinky léků účinky záření MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Accurate enumeration of Paenibacillus mucilaginosus (formerly Bacillus mucilaginosus) bacterium from environmental samples on solid medium is challenging owing to its extensive extracellular polysaccharides (EPS) excretion. In the present study, P. mucilaginosus enumeration has been facilitated by a simple modification: addition of triphenyl tetrazolium chloride (TTC) to growth medium and application of a second soft agar layer. Results show distinctively better and accurate colonies' count. This method can be applied to all bacterial species that produce excessive EPS that may interfere with direct count.
The present paper is focused on zinc(ii) treatment effects on prostatic cell lines PC-3 (tumour) and PNT1A (non-tumour). Oxidative status of cells was monitored by evaluation of expression of metallothionein (MT) isoforms 1A and 2A at the mRNA and protein level, glutathione (oxidised and reduced), and intracellular zinc(ii) after exposition to zinc(ii) treatment at concentrations of 0-150 μM using electrochemical methods, western blotting and fluorescent microscopy. A novel real-time impedance-based growth monitoring system was compared with widely used end-point MTT assay. Impedance-based IC(50) for zinc(ii) is 55.5 and 150.8 μM for PC-3 and PNT1A, respectively. MTT-determined IC(50) are >1.3-fold higher. Impedance-based viability correlates with viable count (r > 0.92; p < 0.03), not with MTT. Two-fold lower intracellular zinc(ii) in the tumour PC-3 cell line was found. After zinc(ii) treatment >2.6-fold increase of intracellular zinc(ii) was observed in non-tumour PNT1A and in tumour PC-3 cells. In PC-3 cells, free and bound zinc(ii) levels were enhanced more markedly as compared to PNT1A. PNT1A produced 4.2-fold less MT compared to PC3. PNT1A cells showed a 4.8-fold increase trend (r = 0.94; p = 0.005); PC-3 did show a significant trend at MT1 and MT2 protein levels (r = 0.93; p = 0.02) with nearly ten-fold increase after 100 μM zinc(ii) treatment. In terms of redox state, PNT1A had a predominance of reduced GSH forms (GSH : GSSG ratio > 1), when exposed to zinc(ii) compared to PC3, where predominance of oxidised forms remains at all concentrations. IC(50) differs significantly when determined by MTT and real-time impedance-based assays due to dependence of impedance on cell morphology and adhesion. When real-time growth monitoring, precise electrochemical methods and fluorescent microscopy are performed together, accurate information for metal fluxes, their buffering by thiol compounds and monitoring of the redox state become a powerful tool for understanding the role of oxidative stress in carcinogenesis.
- MeSH
- buňky - růstové procesy účinky léků MeSH
- fluorescenční barviva chemie MeSH
- fluorescenční mikroskopie metody MeSH
- formazany chemie MeSH
- glutathion metabolismus MeSH
- impedanční spektroskopie MeSH
- lidé MeSH
- lineární modely MeSH
- metalothionein genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory prostaty farmakoterapie metabolismus patologie MeSH
- oxidace-redukce MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- RNA chemie genetika MeSH
- síran zinečnatý farmakologie MeSH
- tetrazoliové soli chemie MeSH
- viabilita buněk fyziologie MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
