L-band Electron Paramagnetic Resonance Spectroscopy Research Articles

In Vivo Electron Paramagnetic Resonance Molecular Profiling of Tumor Microenvironment upon Tumor Progression to Malignancy in an Animal Model of Breast Cancer.

Hypoxia and acidosis are recognized tumor microenvironment (TME) biomarkers of cancer progression. Alterations in cancer redox status and metabolism are also associated with elevated levels of intracellular glutathione (GSH) and interstitial inorganic phosphate (Pi). This study aims to evaluate the capability of these biomarkers to discriminate between stages and inform on a switch to malignancy. These studies were performed using MMTV-PyMT( +) female transgenic mice that spontaneously develop breast cancer and emulate human tumor staging. In vivo assessment of oxygen concentration (pO2), extracellular acidity (pHe), Pi, and GSH was performed using L-band electron paramagnetic resonance spectroscopy and multifunctional trityl and GSH-sensitive nitroxide probes. Profiling of the TME showed significant deviation of measured biomarkers upon tumor progression from pre-malignancy (pre-S4) to the malignant stage (S4). For the combined marker, HOP: (pHe × pO2)/Pi, a value > 186 indicated that the tumors were pre-malignant in 85% of the mammary glands analyzed, and when < 186, they were malignant 42% of the time. For GSH, a value < 3mM indicated that the tumors were pre-malignant 74% of the time, and when > 3mM, they were malignant 80% of the time. The only marker that markedly deviated as early as stage 1 (S1) from its value in pre-S1 was elevated Pi, followed by a decrease of pHe and pO2 and increase in GSH at later stages. Molecular TME profiling informs on alteration of tumor redox and metabolism during tumor staging. Early elevation of interstitial Pi at S1 may reflect tumor metabolic alterations that demand elevated phosphorus supply in accordance with the high rate growth hypothesis. These metabolic changes are supported by the following decrease of pHe due to a high tumor reliance on glycolysis and increase of intracellular GSH, a major intracellular redox buffer. The appreciable decrease in TME pO2 was observed only at malignant S4, apparently as a consequence of tumor mass growth and corresponding decrease in perfusion efficacy and increase in oxygen consumption as the tumor cells proliferate.

Microencapsulation of carbon particles used as oxygen sensors in EPR oximetry to stabilize their responsiveness to oxygen in vitro and in vivo

The electron paramagnetic resonance (EPR) spectra of some paramagnetic materials exhibit a pO2 (partial pressure of oxygen)-dependent linewidth. By recording the EPR linewidth in vivo using low-frequency EPR spectrometers, it is possible to measure the partial pressure of oxygen in tissues. It has been found, however, that some of the paramagnetic materials with optimal spectroscopic properties in vitro may lose or change their responsiveness to oxygen in tissues. The aim of this study was to microencapsulate paramagnetic particles by biopolymers in order to stabilize their responsiveness to oxygen. Carbohydrate char particles (Bubinga) were encapsulated with different biopolymers: cellulose acetate or cellulose triacetate, silicone and polyurethane. The performance of the materials was evaluated in vitro and in vivo. X-band EPR spectroscopy was used to test the variation of the calibration curve (EPR linewidth as a function of the pO2) after incubation in saline and after prolonged residence in tissues. The stability of the responsiveness to pO2 in vivo was carried out by L-band EPR spectroscopy using mice that received injection of the oxygen sensors in the muscles. After residence in saline and prolonged residence in tissues, only the calibration curve of the silicone-coated (coating weight of 0.5% (w/w)) paramagnetic materials remained unchanged, while those of oxygen sensors coated with cellulose acetate, cellulose triacetate and polyurethane changed.