Ferroptosis, a new form of cell death defined after radiation exposure.

PURPOSE: Ferroptosis is an iron-dependent form of regulated cell death, driven by excessive lipid peroxidation and/or inactivation/depletion of protective molecules against lipid peroxidation. Ionizing radiation can induce ferroptosis in both normal tissues and tumor cells. Here, we reviewed the findings of ionizing radiation-induced ferroptosis. CONCLUSIONS: Ionizing radiation induces an increase in hydroxyl radicals, free iron, and lipid metabolic enzymes, which subsequently synergistically initiate a high level of lipid peroxidation, making ionizing radiation an exogenous inducer of ferroptosis. In addition, ferroptosis may be the primary form of cell death in the bone marrow under hematopoietic acute radiation syndrome. Ionizing radiation can also induce changes in iron metabolism, which may be a target for regulating ferroptosis. Finally, ionizing radiation-induced ferroptosis initiates from the cytoplasm and ends on the membrane, and is independent of DNA damage.

Low-Diffusion Fricke Gel Dosimeters with Core-Shell Structure Based on Spatial Confinement.

The diffusion of ferric ions is an important challenge to limit the application of Fricke gel dosimeters in accurate three-dimensional dose verification of modern radiotherapy. In this work, low-diffusion Fricke gel dosimeters, with a core-shell structure based on spatial confinement, were constructed by utilizing microdroplet ultrarapid freezing and coating technology. Polydimethylsiloxane (PDMS), with its excellent hydrophobicity, was coated on the surface of the pellets. The concentration gradient of the ferric ion was realized through shielding half of a Co-60 photon beam field size, and ion diffusion was measured by both ultraviolet-visible spectrophotometry and magnetic resonance imaging. No diffusion occurred between the core-shell pellets, even at 96 h after irradiation, and the diffusion length at the irradiation boundary was limited to the diameter (2-3 mm) of the pellets. Furthermore, Monte Carlo calculations were conducted to study dosimetric properties of the core-shell dosimeter, which indicated that a PDMS shell hardly affected the performance of the dosimeter.

Hematopoietic protection and mechanisms of ferrostatin-1 on hematopoietic acute radiation syndrome of mice.

PURPOSE: Baicalein (an anti-ferroptosis drug) was recently reported to synergistically improve the survival rate of mice following a high dose of total body irradiation with anti-apoptosis and anti-necroptosis drugs. At the same time, our group has demonstrated that ferrostatin-1, a ferroptosis inhibitor, improves the survival rate of a mouse model of hematopoietic acute radiation syndrome to 60% for 150 days (p < .001). These phenomena suggest that ferroptosis inhibition can mitigate radiation damage. In this study, we continued to study the mechanisms by which ferrostatin-1 alleviated radiation-induced ferroptosis and subsequent hematopoietic acute radiation syndrome. MATERIALS AND METHODS: Male ICR mice (8-10 weeks old) were exposed to doses of 0, 8, or 10 Gy irradiated from a 137Cs source. Ferrostatin-1 was intraperitoneally injected into mice 72 h post-irradiation. Bone marrow mononuclear cells (BMMCs) and peripheral blood cells were counted. The changes in iron-related parameters, lipid metabolic enzymes, lipid peroxidation repair molecules (glutathione peroxidase 4, glutathione, and coenzyme Q10), and inflammatory factors (TNF-α, IL-6, and IL-1ß) were evaluated using biochemical or antibody techniques. RESULTS: Ferrostatin-1 increased the number of red and white blood cells, lymphocytes, and monocytes in the peripheral blood after total body irradiation in mice by mitigating the ferroptosis of BMMCs. Total body irradiation induced ferroptosis in BMMCs by increasing the iron and lipid peroxidation levels and depleting the acyl-CoA synthetase long-chain family member 4 (ASCL4), lipoxygenase 15, glutathione peroxidase 4, and glutathione levels. Ferroptotic BMMCs did not release TNF-α, IL-6, or IL-1ß at the early stage of radiation exposure. Ferrostatin-1 mitigated the lipid peroxidation of radiation-induced ferroptosis by attenuating increases in levels of hemosiderin and liable iron pool and decreases in levels of ASCL4 and glutathione peroxidase 4. CONCLUSIONS: The onset of total body irradiation-induced ferroptosis in BMMCs involved changes in iron, lipid metabolic enzymes, and anti-lipid peroxidation molecules. Ferrostatin-1 could be a potential radiation mitigation agent by acting on these targets.

Highly Sensitive Gold Nanoparticles-DNA Nanosensor for γ-Radiation Detection.

It is very important to control the ionizing radiation dose in radiation therapy, which depends on the accurate and rapid measurement of radiation. Herein, a novel and highly sensitive nanosensor for γ-radiation detection is constructed using single-stranded DNA sequences as radiation-sensitive material and gold nanoparticles (AuNPs) as a signal reporter. Well-dispersed AuNPs gradually aggregated at high salt concentration when the sensor was irradiated, and this change was quantified by the visible spectra and surface plasmon resonance spectra. The radiation nanosensor has excellent linearity in the dose range of 0-100 Gy under optimal conditions. This method is simple and fast, which provides a new path for the γ-radiation dosimeter and has potential applications in the assessment of radiation-induced biological effects.

Ionizing radiation induces ferroptosis in granulocyte-macrophage hematopoietic progenitor cells of murine bone marrow.

Purpose: To study whether radiation-induced bleeding in the bone marrow induced iron accumulation, and subsequently caused ferroptosis in granulocyte-macrophage hematopoietic progenitor cells.Materials and methods: Male mice were subjected to different doses (0, 4, 8, or 10 Gy) of gamma radiation from a 137Cs source. The changes in iron metabolism or ferroptosis-related parameters of irradiated bone marrow were accessed with biochemical, histopathological, and antibody methods. Hematocytes were detected with a hematology analyzer. The counts of granulocyte-macrophage hematopoietic progenitor cells were measured with the granulocyte-macrophage colony-forming unit.Results: Iron accumulation occurred in the bone marrow, which caused by radiation-induced hemorrhage. The iron accumulation triggered an iron regulatory protein-ferroportin 1 axis to increase serum iron levels. Using LDN193189, radiation-induced iron accumulation was demonstrated to decrease white blood cell counts at least partly through a decrease in the counts of granulocyte-macrophage hematopoietic progenitor cells. The reduction in the counts of granulocyte-macrophage hematopoietic progenitor cells was subsequently demonstrated to attribute to ferroptosis with the use of ferroptosis inhibitors and through the detection of ferroptosis related-parameters. The survival rate of irradiated mice was improved using Ferrostatin-1 or LDN193189.Conclusions: These findings suggest that radiation-induced hemorrhage in the bone marrow causes ferroptosis in granulocyte-macrophage hematopoietic progenitor cells, and anti-ferroptosis has the potential to be a radioprotective strategy to ameliorate radiation-induced hematopoietic injury.

Evaluation of the Effect of a Tracheal Stent on Radiation Dose Distribution via Micro-CT Imaging.

PURPOSE: To study the effect of a metal tracheal stent on radiation dose distribution. METHOD: A metal tube bracket is placed in a self-made foam tube sleeve, and micro-computed tomography scanning is performed directly. The foam sleeve containing the metal bracket is placed in a nonuniform phantom for a routine computed tomography scan. The stents in conventional computed tomography images are replaced by the stents in micro-computed tomography images. Subsequently, 2 sets of computed tomography images are obtained and then imported to a radiotherapy treatment planning system. A single photon beam at 0° is designed in a field size of 10 cm × 10 cm, a photon beam of 6 MV, and a monitor unit of 200 MU. Monte Carlo algorithm is used to calculate the dose distribution and obtain the dose curve of the central axis of the field. The dose is verified with thermoluminescence dose tablets. RESULTS: The micro-computed tomography images of the tracheal stent are clearer and less false-like than its conventional computed tomography images. The planned dose curves of the 2 groups are similar. In comparison with the images without any stents in place, the doses at the incident surface of the stent in the conventional computed tomography images and at the stent exit surface in the rear of the stent increase by 1.86% and 2.76%, respectively. In the micro-computed tomography images, the doses at the incident surface of the stent and at the exit surface behind the stent increase by 1.32% and 1.19%, respectively. Conventional computed tomography reveals a large deviation between the measured and calculated values. CONCLUSION: Tracheal stent based on micro-computed tomography imaging has a less effect on radiotherapy calculation than that based on conventional computed tomography imaging.

γ-Radiation Enhanced Luminescence of Thiol-Capped Quantum Dots in Aqueous Solution.

Quantum dots (QDs) have attracted great attention due to their unique optical properties. High fluorescence efficiency is very important for their practical application. In this study, we report a simple and efficient strategy to enhance the photoluminescence of water-dispersed thiol-capped QDs using γ-radiation. Three kinds of QDs with different surface ligands and cores (MPA-CdTe, MPA-CdSe and Cys-CdTe) were fabricated and irradiated by high-energy γ-ray in an aqueous solution. Their photoluminescence intensities were significantly enhanced after irradiation, which were closely related to the radiation dose and the structure of QDs. The positions of the fluorescence emission peaks did not shift obviously after irradiation. The mechanism of photoluminescence enhancement was discussed based on the results of photoluminescence (PL) spectra, UV-visible light absorption (UV-vis) spectra, transmission electron microscope (TEM), X-ray diffraction (XRD) patterns, Fourier transform infrared (FT-IR) spectra and X-ray photoelectron spectroscopy (XPS). This method can be employed to uniformly treat large batches of QDs at room temperature and without other chemicals.

Mechanisms of strontium's adsorption by Saccharomyces cerevisiae: Contribution of surface and intracellular uptakes.

The objective of this work was to explore the mechanisms participating in strontium sorption by living Saccharomyces cerevisiae (S. cerevisiae). The location of strontium adsorbed by S. cerevisiae was studied by our plasmolysis treatment. The contribution of physical and chemical mechanisms was determined quantitatively by desorption and blockage of functional groups. Moreover, our results indicated that bioaccumulation also played a major role in biosorption by living cells. Thus, supplementary methods including 2-DE (two-dimensional electrophoresis) and Matrix-Assisted Laser Desorption/Ionization Tandem Time of Flight Mass Spectrometry (MALDI-TOF-TOF) were employed to analyze the different proteins. The subsequent desorption % of Sr2+ by Distilled Water (DW), NH4NO3 and EDTA-Na2 from Sr2+ loaded sorbents indicated a minor role for physical adsorption, while ion exchange and complexation were responsible for approximately 20% and 40%. Specific blockage of functional groups revealed that carboxyl and amine groups played an important role in Sr2+ binding to the living S. cerevisiae. From our MALDI-TOF-TOF results, we concluded that 38 proteins showed up-regulated expression profiles and 11 proteins showed down-regulated after biosorption. Moreover, proteins belong to: phagocytic function (Act1p); ion channel (S-adenosylmethionine synthase); glycolysis (Tubulin) may directly involve in strontium bioaccumulation. In conclusion, the present work indicates that the strontium sorption mechanism by living S. cerevisiae is complicated including ion-exchange along with complexation as the main mechanism, whereas the other mechanisms such as physical adsorption play a minor contribution. Metabolically-dependent proteins may play an important role in bioaccumulation.

Enhancement of radiotherapy efficacy by silver nanoparticles in hypoxic glioma cells.

Radiotherapy is one of the most widely used treatments for therapy of malignant tumors, but resistance to radiation of hypoxic cells in tumor tissues is still a serious concern. Previous studies have demonstrated that silver nanoparticles (AgNPs) enhance the radiosensitivity of human glioma cells in vitro, but the effect of AgNPs on hypoxic glioma cells has not been investigated in detail. The main purpose of this study is to evaluate the radiosensitizing efficacy of AgNPs on hypoxic glioma cells. The half maximal inhibitory concentration (IC50) values of AgNPs for the hypoxic U251 cells and C6 cells were 30.32 µg/mL and 27.53 µg/mL, respectively. The sensitization enhancement ratio (SER) demonstrated that AgNPs exhibit higher capacity in radiosensitization in hypoxic cells (U251: 1.78; C6: 1.84) than that in normoxic cells (U251: 1.34; C6: 1.45). The underlying mechanism of AgNPs' radiosensitization in hypoxic cells is through the promotion of apoptosis and enhanced destructive autophagy. There is evidence of crosstalk between apoptosis and autophagy in AgNPs-radiosensitized hypoxic cells where inhibition of autophagy results in decreased apoptosis. These findings suggest that AgNPs can be used as a highly effective nano-radiosensitizer for the treatment of hypoxic glioma.

Radiosensitivity enhancement of Fe3O4@Ag nanoparticles on human glioblastoma cells.

Radiotherapy is one of the main therapeutic methods for cancers, but radiation resistance of cancer cells still remains a serious concern. Searching for radiosensitizers to overcome such resistance is therefore urgently required. The goal of this study is to evaluate and compare the radiosensitizing efficacy of Fe3O4-OA, Ag and Fe3O4@Ag nanoparticles on U251 cells. The results show that Fe3O4@Ag nanoparticles have the highest ability of radiosensitization among the three nanoparticles. The underlying mechanism of Fe3O4@Ag nanoparticles' radiosensitivity enhancement is through decrease of the cytoprotective autophagy at the early stage, and increase of the calcium-dependent apoptosis at the later stage. These findings suggest the potential application of Fe3O4@Ag nanoparticles as a highly effective nano-radiosensitizer for the treatment of glioblastoma cells.

UV-enhanced cytotoxicity of CdTe quantum dots in PANC-1 cells depend on their size distribution and surface modification.

The cytotoxicity of quantum dots (QDs) under normal conditions has received more and more attention, but their cytotoxicity under light illumination has not been fully investigated. In this study, different sized CdTe QDs coated with mercaptopropionic acid (MPA) and N-acetylcysteine (NAC) were employed to investigate the influences of size distribution and surface modification on their UV-enhanced cytotoxicity and mechanism. The results indicated that different sized MPA-CdTe QDs exhibited distinct cytotoxicity under UV illumination and the smaller-sized QDs presented more obviously damages to cells than the larger-sized QDs. Comparing with MPA-CdTe QDs, NAC-CdTe QDs had better cellular metabolizability and lower cytotoxicity. The generation of reactive oxygen species (ROS) were also investigated. The results revealed that ROS in cells containing MPA-CdTe QD538 were about 1.7 times of NAC-CdTe QD538 under UV illumination. ROS might play an important role in the UV-enhanced cytotoxicity of QDs. By selecting appropriate surface modifications and particle sizes, the cytotoxicity of QDs under UV illumination could be controlled.

The combined influence of surface modification, size distribution, and interaction time on the cytotoxicity of CdTe quantum dots in PANC-1 cells.

Mercaptopropionic acid (MPA) and cysteamine (Cys) capped CdTe quantum dots (QDs) were successfully prepared and used to investigate the combined influence of surface modification, size distribution, and interaction time on their cytotoxicity in human pancreatic carcinoma (PANC-1) cells. Results indicated that the smaller the size of MPA-CdTe QDs, the higher the cytotoxicity, which could be partly due to the difference of their distribution inside cells. Comparing with MPA-CdTe QDs, Cys-CdTe QDs had better cellular metabolizability and lower cytotoxicity. These QDs' cellular distribution and cytotoxicity were closely related to their interaction time with cells. Their cytotoxicity was found to be significantly enhanced with the increase of incubation time in medium. After QD treatments, the influence of recover time on the final cell viability was also dependent on the concentration and surface modification of QDs used in pretreatment. The combined influence of these factors discussed here might provide useful information for understanding and reducing the cytotoxicity of QDs in future biomedical applications.

Subcellular tracking of drug release from carbon nanotube vehicles in living cells.

The direct observation of drug release from carbon nanotube vehicles in living cells is realized through a unique two-dye labeling approach. Single-walled carbon nanotubes (SWNTs) are firstly marked with fluorescein isothiocyanate (FITC) to track their location and movement inside the cell. Then a fluorescent anticancer drug doxorubicin (DOX) is attached by means of π-stacking onto SWNTs. Delivered by SWNTs into cells, DOX will detach from the vehicle in an acidic environment due to the pH-dependent π-π stacking interaction between DOX and SWNTs. From observation of the two different kinds of fluorescence (green and red) that respectively represent the carrier SWNTs and drug DOX, the process of drug release inside the living cell can be monitored under a confocal microscope. Results show that the drug DOX detaches from SWNTs inside the lysosomes to yield free molecules and escape into the cytoplasm and finally into the nucleus, while the vehicle SWNTs are trapped inside the lysosomes, without entering the nucleus. The current observations confirm previously proposed mechanisms for drug/DOX release inside cells. The experimental establishment of drug-release mechanisms in living cells here might provide important insights for future design of new drug-delivery and release systems.

One-step fabrication of biocompatible chitosan-coated ZnS and ZnS:Mn2+ quantum dots via a γ-radiation route.

Biocompatible chitosan-coated ZnS quantum dots [CS-ZnS QDs] and chitosan-coated ZnS:Mn2+ quantum dots [CS-ZnS:Mn2+ QDs] were successfully fabricated via a convenient one-step γ-radiation route. The as-obtained QDs were around 5 nm in diameter with excellent water-solubility. These QDs emitting strong visible blue or orange light under UV excitation were successfully used as labels for PANC-1 cells. The cell experiments revealed that CS-ZnS and CS-ZnS:Mn2+ QDs showed low cytotoxicity and good biocompatibility, which offered possibilities for further biomedical applications. Moreover, this convenient synthesis strategy could be extended to fabricate other nanoparticles coated with chitosan.PACS: 81.07.Ta; 78.67.Hc; 82.35.Np; 87.85.Rs.

Gamma-radiation synthesis of silk fibroin coated CdSe quantum dots and their biocompatibility and photostability in living cells.

Silk fibroin coated CdSe quantum dots (SF-CdSe QDs) were successfully synthesized via a one-step gamma-radiation route in an aqueous system at room temperature. The as prepared products were characterized by transmission electron microscope (TEM), energy dispersion spectrum (EDS), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectrometer (FT-IR), X-ray diffraction (XRD), ultraviolet-visible spectroscopy (UV-vis) and photoluminescence spectrum (PL). The SF-CdSe QDs were about 5 nm in diameter and exhibited excellent water-solubility and photoluminescence properties. The cellular distribution, photostability and cytotoxicity of SF-CdSe QDs with different amount of SF coatings were also investigated by laser scanning confocal microscope (LSCM) and MTT assays in human pancreatic carcinoma (PANC-1) cells. All the results reveal that these QDs could be easily internalized by cells and localized in cytoplasm around nuclei. Moreover, SF-CdSe QDs were proved to be low cytotoxicity (the concentration of QDs < 5 microg mL(-1)) and high photostability (the illumination energy density < 2 x 10(-5) W microm(-2)) within PANC-1 cells, which was mainly due to the biocompatible silk fibroin. The resulted SF-CdSe QDs might have many potential applications in tumor imaging and therapy. And the synthesis strategy could be easily extended to fabrication of other nanoparticles coated with silk fibroin.

UV-enhanced cytotoxicity of thiol-capped CdTe quantum dots in human pancreatic carcinoma cells.

Quantum dots (QDs) have been gaining popularity due to their potential application in cellular imaging and diagnosis, but their cytotoxicity under light illumination has not been fully investigated. In this study, green and red mercaptopropionic acid capped CdTe quantum dots (MPA-CdTe QDs) were employed to investigate their cytotoxicity in human pancreatic carcinoma cells (PANC-1) under UV illumination. MPA-CdTe QDs exhibited excellent photostability under UV illumination and could be easily ingested by cells. The cytotoxicity of MPA-CdTe QDs was significantly enhanced under UV illumination, which was determined by changes in cell morphology as well as by decreases in the metabolic activity and cell counting. Our results indicated that green and red QDs had different cellular distribution and exhibited distinct UV-enhanced cytotoxicity. UV illumination enhanced the generation of reactive oxygen species (ROS) in cells containing QDs, and NAC antioxidant could reduce their damage to cells under UV illumination. Moreover, the influences of different UV illumination conditions on the viability of cells containing QDs were examined and discussed in detail.

Cancer-cell targeting and photoacoustic therapy using carbon nanotubes as "bomb" agents.

A unique approach using the large photoacoustic effect of single-walled carbon nanotubes (SWNTs) for targeting and selective destruction of cancer cells is demonstrated. SWNTs exhibit a large photoacoustic effect in suspension under the irradiation of a 1064-nm Q-switched millisecond pulsed laser and trigger a firecracker-like explosion at the nanoscale. By using such an explosion, a photoacoustic agent is developed by functionalizing the SWNTs with folate acid (FA) that can selectively bind to cancer cells overexpressing folate receptor on the surface of the cell membrane and kill them through SWNT explosion inside the cells under the excitation of millisecond pulsed laser. The uptake pathway of folate-conjugated SWNTs into cancer cells is investigated via fluorescence imaging and it is found that the FA-SWNTs can enter into cancer cells selectively with a high targeting capability of 17-28. Under the treatment of 1064-nm millisecond pulsed laser, 85% of cancer cells with SWNT uptake die within 20 s, while 90% of the normal cells remain alive due to the lack of SWNTs inside cells. Temperature changes during laser treatment are monitored and no temperature increases of more than +/- 3 degrees C are observed. With this approach, the laser power used for cancer killing is reduced 150-1500 times and the therapy efficiency is improved. The death mechanism of cancer cells caused by the photoacoustic explosion of SWNTs is also studied and discussed in detail. These discoveries provide a new way to use the photoacoustic properties of SWNTs for therapeutic applications.

Gamma-radiation synthesis and properties of superparamagnetic CS-ZnSe:Mn nanocrystals for biological labeling.

Chitosan coated ZnSe:Mn (CS-ZnSe:Mn) nanocrystals were successfully synthesized in aqueous system through a gamma-radiation route at room temperature under ambient pressure. The structure and properties of nanocrystals were investigated with transmission electron microscope (TEM), fourier transform infrared spectrometer (FT-IR), ultraviolet-visible (UV-vis) spectrometer, photoluminescence emission (PL) spectra, X-ray Diffraction (XRD) and energy dispersion spectrum (EDS). Results showed that the diameter of these nanocrystals was about 4 nm with narrow size distribution. With the increase of doped Mn2+ concentration, strong emission peak at 610 nm was observed besides the weak emission peak at 425 nm since the non-radiative transition of 4T1(4G)-6A1(6S) level, resulting the transfer of fluorescence color from blue to orange. Moreover, analysis of SQUID magnetometer indicated that the nanocrystals were superparamagnetic with a saturation magnetization of 1.7 emu/g and a Curie-Weiss temperature of 14-15 K. Hep G2 cells were incubated in solution of nanocrystals and results showed that the synthesized nanocrystals could stain cytoplasm but could not enter into nucleus.