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1.
Nanoscale ; 8(37): 16710-16718, 2016 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-27714168

RESUMO

Use of multicellular tumor spheroids (MTS) to investigate therapies has gained impetus because they have potential to mimic factors including zonation, hypoxia and drug-resistance. However, analysis remains difficult and often destroys 3D integrity. Here we report an optical technique using targeted nanosensors that allows in situ 3D mapping of redox potential gradients whilst retaining MTS morphology and function. The magnitude of the redox potential gradient can be quantified as a free energy difference (ΔG) and used as a measurement of MTS viability. We found that by delivering different doses of radiotherapy to MTS we could correlate loss of ΔG with increasing therapeutic dose. In addition, we found that resistance to drug therapy was indicated by an increase in ΔG. This robust and reproducible technique allows interrogation of an in vitro tumor-model's bioenergetic response to therapy, indicating its potential as a tool for therapy development.


Assuntos
Nanoestruturas , Neoplasias/química , Análise Espectral Raman , Esferoides Celulares/química , Humanos , Concentração de Íons de Hidrogênio , Células MCF-7 , Oxirredução , Microambiente Tumoral
2.
Faraday Discuss ; 187: 501-20, 2016 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-27032696

RESUMO

Measuring markers of stress such as pH and redox potential are important when studying toxicology in in vitro models because they are markers of oxidative stress, apoptosis and viability. While surface enhanced Raman spectroscopy is ideally suited to the measurement of redox potential and pH in live cells, the time-intensive nature and perceived difficulty in signal analysis and interpretation can be a barrier to its broad uptake by the biological community. In this paper we detail the development of signal processing and analysis algorithms that allow SERS spectra to be automatically processed so that the output of the processing is a pH or redox potential value. By automating signal processing we were able to carry out a comparative evaluation of the toxicology of silver and zinc oxide nanoparticles and correlate our findings with qPCR analysis. The combination of these two analytical techniques sheds light on the differences in toxicology between these two materials from the perspective of oxidative stress.


Assuntos
Nanopartículas Metálicas/toxicidade , Análise Espectral Raman/métodos , Testes de Toxicidade/métodos , Algoritmos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Prata/toxicidade , Óxido de Zinco/toxicidade
3.
Biochem Soc Trans ; 42(4): 899-904, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25109976

RESUMO

The measurement of intracellular analytes has been key in understanding cellular processes and function, and the use of biological nanosensors has revealed the spatial and temporal variation in their concentrations. In particular, ratiometric nanosensors allow quantitative measurements of analyte concentrations. The present review focuses on the recent advances in ratiometric intracellular biological nanosensors, with an emphasis on their utility in measuring analytes that are important in cell function.


Assuntos
Nanotecnologia/instrumentação , Espectrometria de Fluorescência , Análise Espectral Raman
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