Nanoscopic synchrotron X-ray imaging was performed on scalp hair samples of patients with breast cancer and healthy individuals to investigate any structural differences as diagnostic tool. Hair strands were divided into 2-3 segments along the strands from root to tip, followed by imaging either in projection or in CT scanning with a monochromatic 6.78-keV X-ray using zone-plate optics with a resolving power of 60 nm. All the examined cancer hairs exhibited medulla loss with cancer stage-dependent pattern; complete loss, discontinuous or trace along the strands. In contrast, medullas were well retained without complete loss in the healthy hair. In the CT-scanned axial images, the cortical spindle compartments had no contrast in the healthy hair, but appeared hypointense in contrast to the surrounding hyperintense cortical membrane complex in the cancer hair. In conclusion, observation of medulla loss and cortical membrane enhancements in the hair strands of breast cancer patients demonstrated structural variations in the cancer hair, providing a new platform for further synchrotron X-ray imaging study of screening breast cancer patients.
Bursts of emissions of low-energy electrons, including interatomic Coulomb decay electrons and Auger electrons (0-1000 eV), as well as X-ray fluorescence produced by irradiation of large-Z element nanoparticles by either X-ray photons or high-energy ion beams, is referred to as the nanoradiator effect. In therapeutic applications, this effect can damage pathological tissues that selectively take up the nanoparticles. Herein, a new nanoradiator dosimetry method is presented that uses probes for reactive oxygen species (ROS) incorporated into three-dimensional gels, on which macrophages containing iron oxide nanoparticles (IONs) are attached. This method, together with site-specific irradiation of the intracellular nanoparticles from a microbeam of polychromatic synchrotron X-rays (5-14 keV), measures the range and distribution of OH radicals produced by X-ray emission or superoxide anions ({\rm{O}}_2^-) produced by low-energy electrons. The measurements are based on confocal laser scanning of the fluorescence of the hydroxyl radical probe 2-[6-(4'-amino)phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF) or the superoxide probe hydroethidine-dihydroethidium (DHE) that was oxidized by each ROS, enabling tracking of the radiation dose emitted by the nanoradiator. In the range 70 µm below the irradiated cell, ^\bullet{\rm{OH}} radicals derived mostly from either incident X-ray or X-ray fluorescence of ION nanoradiators are distributed along the line of depth direction in ROS gel. In contrast, {\rm{O}}_2^- derived from secondary electron or low-energy electron emission by ION nanoradiators are scattered over the ROS gel. ROS fluorescence due to the ION nanoradiators was observed continuously to a depth of 1.5 mm for both oxidized APF and oxidized DHE with relatively large intensity compared with the fluorescence caused by the ROS produced solely by incident primary X-rays, which was limited to a depth of 600 µm, suggesting dose enhancement as well as more penetration by nanoradiators. In conclusion, the combined use of a synchrotron X-ray microbeam-irradiated three-dimensional ROS gel and confocal laser scanning fluorescence microscopy provides a simple dosimetry method for track analysis of X-ray photoelectric nanoradiator radiation, suggesting extensive cellular damage with dose-enhancement beyond a single cell containing IONs.
The emission of fluorescent X-rays and low-energy electrons by mid-/high-Z nanoparticles upon irradiation with either X-ray photons or high-energy ion beams is referred to as the nanoradiator effect (NRE). A track analysis of NRE was performed using reactive oxygen species (ROS) gels, to which macrophages containing gold nanoparticles (AuNPs) were attached, together with single-cell irradiation of the intracellular nanoparticles from a microbeam of synchrotron X-rays, and the range and distribution of ^\bulletOH and O2^{ \bullet - } produced were compared with those of the Fe-nanoradiator by magnetite nanoparticles (FeONP, Fe3O4). The Au-nanoradiator generated ROS fluorescence to a greater depth and wider angle with respect to the incident X-rays than that of the Fe-nanoradiator. The ROS-oxidant fluorescence intensity ratios of ^\bulletOH to O2^{ \bullet - } were different for the AuNPs and FeONPs, reflecting different relative yields of electrons and fluorescent X-rays from NRE. In the region immediately (<100 µm) below the irradiated cell, ^\bulletOH-radicals were distributed mainly along two or three tracks in the depth direction in the FeONP- or AuNP-ROS gel. In contrast, O2^{ \bullet - } was scattered more abundantly in random directions in the AuNP-ROS gel than in the FeONP-ROS gel. Track analysis of X-ray photoelectric nanoradiator radiation showed a different range of dose distribution and relative emission compositions between Au- and Fe-nanoradiators, suggesting more extensive damage beyond a single cell containing AuNPs than one containing FeONPs.
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