Incorporation of deuterium oxide in MCF-7 cells to shed further mechanistic insights into benzo[a]pyrene-induced low-dose effects discriminated by ATR-FTIR spectroscopy

Abstract

This study evaluated the potential of deuteration to enhance the mechanistic information obtainable by biospectroscopy techniques in biological-cell models. These techniques were previously demonstrated to identify low-dose effects (≤nM) induced by test agents; this is of critical interest in terms of developing novel approaches to monitor environmentally-induced cell alterations. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was coupled with multivariate analysis to characterize a low-dose (10(-10) M) compared to a high-dose (10(-6) M) exposure of benzo[a]pyrene (B[a]P) in oestrogen-responsive MCF-7 cells; these results were used as a positive control for spectroscopic detection of B[a]P-induced effects. Deuterium oxide (D2O) was then applied as part of a fixative solution and/or at low levels incorporated into growth medium prior to ATR-FTIR spectrochemical analysis. The application of D2O as an alternative solvent in spectroscopy is widespread, but D2O has never before been applied to biospectroscopic analysis of in vitro toxicology assays. This allowed comparison between deuterated- and typically-derived IR spectra, facilitating significant insights into the effects of deuteration, and suggested that the addition of D2O to biospectroscopy assays could improve understanding of low-dose effects.

4-Nonylphenol effects on rat testis and sertoli cells determined by spectrochemical techniques coupled with chemometric analysis

“…Therefore, in the present study, both ATR-FTIR and Raman spectroscopy were employed to detect biomolecular alterations in testis from NP-treated rats of different ages and NP-treated sertoli cells in vitro. Following spectroscopic measurements, spectral data were analysed using principal component analysis followed by linear discriminant analysis (PCA-LDA), which generates scores plots in two-or three-dimensional spaces and allows the construction of cluster vectors (Heppenstall et al, 2013) for data visualization. The aim of this study was to determine the NP-induced effects on biomolecular parameters of testis in a dose-and age-related manner.…”