RBE of 10 kV X rays determined for the human mammary epithelial cell line MCF-12A

The dependence of relative biological effectiveness (RBE) on photon energy is a topic of extensive discussions. The increasing amount of in vitro data in the low-energy region indicates this to be a complex dependence that is influenced by the end point and cell line studied. In the present investigation, the RBE of 10 kV X rays (W anode) was determined relative to 200 kV X rays (W anode, 0.5 mm copper filter) for cell survival in the dose range 1-10 Gy and for induction of micronuclei in the range 0.5-3.6 Gy for MCF-12A human mammary epithelial cells. The RBE for cell survival was found to increase with decreasing dose, being 1.21+/-0.03 at 10% survival. Considerably higher values were obtained for micronucleus induction, where the RBE(M) obtained from the ratio of the linear coefficients of the dose-effect curves was 2.6+/-0.4 for the fraction of binucleated cells with micronuclei and 4.1+/-1.0 for the number of micronuclei per binucleated cell. These values, together with our previous data, support a monotonic increase in RBE with decreasing photon energy down to the mean energy of 7.3 keV used in the present study.     

Yield of sister chromatid exchanges and survival of V79-4 Chinese hamster cells irradiated with 0.7 MeV neutrons

The dependence of the survival rate and the number of sister chromatid exchanges (SCEs) in Chinese hamster V79-4 cells on the dose of gamma-rays and neutrons with average energy of 0.7 MeV has been investigated. The value of RBE for neutrons is 5.5. The number of SCEs increased with the dose of gamma-radiation while no induction of SCEs could be detected after neutron irradiation.     

Radiobiology of ultrasoft X rays. IV. Flat and round-shaped hamster cells (CHO-10B, HS-23)

The results reported earlier in this series indicated that the relative biological effectiveness (RBE) of ultrasoft X rays decreases with decreasing cell thickness, approaching unity for the thinnest cells used, plateau-phase human skin fibroblasts (HSF). The possible dependence of RBE on the configuration of the cell nucleus is investigated further in this paper using two CHO cell lines that attach well and have similar intrinsic radiosensitivities to 60Co gamma rays. One of the lines forms monolayers similar to V79 cells, while the other remains more spherical during growth. We find an increasing RBE with decreasing X-ray energy for both of these cell lines, consistent with our results using V79 cells. Also consistent with our results obtained with 10T1/2 and HSF cells, we find an increasing RBE with increasing cell thickness. The possible dependence of RBE on radiosensitivity and the use of the concept of mean dose for ultrasoft X rays is discussed.     

Low-LET microbeam investigation of the track-end dependence of electron-induced damage in normal human diploid fibroblasts

Using a pulsed electron beam, we investigated the dependence of micronucleus formation on the incident electron energy in AG01522 human diploid fibroblasts after nontargeted irradiations at 25 and 80 keV. Examining the dose response, we found that 25 keV electrons are more effective than 80 keV electrons at producing biological damage for a given dose. Our results demonstrating the induction of micronuclei as a function of incident electron energy offer direct support for the hypothesis that the electron track end is responsible for the biological damage occurring in the cell.     

Model of mammalian cell reproductive death II. Comparison with experimental data and discussion

A general equation for mammalian cell survival has been derived in the previous paper. This paper presents the results of comparison of theoretical evaluations with survival data available from the literature, including different cell lines, variations in linear energy transfer, dose rate and dose fractionation effects and the effects of ultrasoft X-rays and superheavy ions. Merits and demerits of the model are considered in comparison with other models of radiation-induced killing of mammalian cells published in the literature.     

Energy and dose dependence of GafChromic EBT3-V3 film across a wide energy range

AimTo determine the energy and dose dependence of GafChromic EBT3-V3 film over an energy range 0.2 mm Al HVL to 6 MV.BackgroundThe decay scheme of a brachytherapy source may be complex and the spectrum of energy can be wide. LiF TLDs are the golden standard recommended for dosimetric measures in brachytherapy, for their energy independence, but TLDs could be not available in some centres. An alternative way to perform dose measurements is to use GafChromic films, but they show energy dependence.Methods and materialsFilms have been irradiated at increasing dose with three different beams: 6 MV beam, TPR20, 10 = (0.684 ± 0.01), HVL = (2.00 ± 0.01)mmAl and HVL = (0.20 ± 0.01)mmAl. Calibration curves were generated using the same dose range (0cGy to 850cGy) for the three energies. Using the 6 MV calibration curve as reference, the film response in terms of net optical density (OD) was evaluated.ResultsThe difference in the calibration curve obtained by irradiating the film with 6 MV and 2 mm Al HVL energy beams is less than 3 %, within the calibration uncertainty, in the dose range 500-850cGy. The OD of EBT3-V3 film is significantly lower at 0.2 mmAl HVL compared to 6 MV, showing differences up to 25 %.ConclusionWithin the range 6 MV-2 mm Al HVL and dose higher than 500cGy, GafChromic EBT3-V3 films are energy independent. In this dose range, films can be calibrated in a simple geometry, using a 6 MV Linac beam, and can be used for brachytherapy sources dose measures. The use of EBT3 films can be extended to reference dosimetry in Ir-192 clinical brachytherapy.     

Dosimetric uncertainties impact on cell survival curve with low energy proton

There is an increasing number of radiobiological experiments being conducted with low energy protons (less than 5 MeV) for radiobiological studies due to availability of sub-millimetre focused beam. However, low energy proton has broad microdosimetric spectra which can introduce dosimetric uncertainty. In this work, we quantify the impact of this dosimetric uncertainties on the cell survival curve and how it affects the estimation of the alpha and beta parameters in the LQ formalism. Monte Carlo simulation is used to generate the microdosimetric spectra in a micrometer-sized water sphere under proton irradiation. This is modelled using radiobiological experiment set-up at the Centre of Ion Beam Application (CIBA) in National University of Singapore. Our results show that the microdosimetric spectra can introduce both systematic and random shifts in dose and cell survival; this effect is most pronounced with low energy protons. The alpha and beta uncertainties can be up to 10% and above 30%, respectively for low energy protons passing through thin cell target (about 10 microns). These uncertainties are non-negligible and show that care must be taken in using the cell survival curve and its derived parameters for radiobiological models.     

Tumor control probability model for alpha-particle-emitting radionuclides

Alpha-particle emitters are currently being evaluated for the treatment of metastatic disease. The dosimetry of alpha-particle emitters is a challenge, however, because the stochastic patterns of energy deposition within cellular targets must be taken into account. We propose a model for the tumor control probability of alpha-particle emitters which takes into account these stochastic effects. An expression for cell survival, which is a function of the microdosimetric single-event specific-energy distribution, is multiplied by the number of cells within the tumor cluster. Poisson statistics is used to model the probability of zero surviving cells within the cluster. Based on this analysis, a number of observations have been made: (1) The dose required to eradicate a tumor is nearly a linear function of the cell survival parameter z(0). (2) Cells with smaller nuclei will require more dose to achieve the same level of tumor control probability, relative to cells with larger nuclei, for an identical source-target configuration and cell sensitivity. (3) As the targeting of alpha-particle emitters becomes more specific, the dose required to achieve a given level of tumor control decreases. (4) Additional secondary effects include cell shape and the initial alpha-particle energy.     

A systemic model of the reproductive death of mammalian cells. The effect of radiation dose rate and fractionation

On the basis of the previously developed systemic model a study was made of the effect of dose rate on the survival of mammalian cells, RBE of small doses of heavy ions, and fractionation of radiation. There was a good agreement between theoretical and experimental results. The calculations showed that D10 (10% survival dose) is a function of dose rate P even for such ions as helium and boron which, however, exhibited an insignificant dependence of D10 on P (within the range from (10(-1) to 1 cGy/min). The influence of repair and the rate of cell division on RBE of radiation was determined.     

Simulation of binary methods for the microdosimetric analysis of cell survival after alpha-particle irradiation: ability to distinguish between different models

Analysis of cell survival after alpha-particle irradiation must account for the distribution in the amounts of energy deposited in each cell nucleus. Microdosimetric computations are usually used to determine these distributions. Irradiation with microbeams and other modern techniques has made these computations unnecessary for certain cell geometries. These techniques allow the survival of individual cells to be correlated with the amount of radiation delivered to individual cell nuclei. However, to maintain the individuality of data generated for each cell, new methods of analysis are required. In this study, we propose the use of binary methods. Each cell is regarded as a Bernoulli trial with a different probability for success (colony formation). Parameter values of the survival model are chosen to maximize the likelihood of the observed outcome. To evaluate this method, simulated data for 500, 5000 and 50,000 cells irradiated by alpha particles are analyzed along with the associated outcome for four different cell survival models. Each survival model has a different dependence on the radius of the cell nucleus. These results indicate that the model that was simulated has the highest likelihood value in all cases. However, the ability to distinguish between competing models is present only for a larger numbers of cells.     

The influence of dose rate on the lethal and mutagenic effects of X-rays in proliferating L5178Y cells differing in radiation sensitivity

The lethal and mutagenic effects of ionizing radiation delivered at high (53 Gy/h) and low (0.02 Gy/h) dose rates were measured in two closely related strains of mouse lymphoma L5178Y cells differing in radiation sensitivity (LY-R and LY-S). Strain LY-R was more resistant to the lethal effects of radiation than strain LY-S when exposed at either the high or low dose rate. The survival of strain LY-R was markedly enhanced by the reduction in dose rate. The dose-rate dependence of the survival of strain LY-S was less clear, because of the biphasic nature of its survival curve following low dose-rate radiation. However, if the initial slope of the low dose-rate survival curve is compared to the slope of the high dose-rate survival curve for strain LY-S, only a slight increase in survival at the low dose rate is apparent. Although more sensitive to the lethal effects of radiation, strain LY-S was less mutable at the hypoxanthine/guanine phosphoribosyl transferase locus by both low dose-rate and high dose-rate radiation than strain LY-R. Little dose-rate dependence was exhibited by either strain with regard to the mutagenic effects of radiation. Thus, for strain LY-R, which showed marked dose-rate dependence for survival but not for mutation, the ratio of mutational to lethal lesions was much greater following exposure to low dose-rate than to high dose-rate radiation.     

The radiation dose to cells in vitro from intracellular indium-111

Most of the radionuclides used in nuclear medicine emit low energy Auger electrons following radioactive decay. These emissions, if intracellular, could irreparably damage the radiosensitive structures of the cell. The resulting radiation dose, which is a measure of biological damage in the affected cell, could be many times the average radiation dose to the associated organ. In this series of experiments, the radiation dose to the nucleus of a chinese hamster V79 cell was determined for the intracellular radiopharmaceutical 111indium-oxine. Assuming the cell nucleus to be the radiosensitive volume, the radiation dose would be primarily due to the low energy Auger electrons. A much smaller dose would be absorbed from the penetrating X- and gamma-rays and internal conversion electrons released from other radiolabelled cells in the culture. The radiation dose to the cell from the intranuclear decay of 111In was empirically established from cell survival studies to be 3.5 mGy/decay, using cobalt-60 as a reference radiation. The average dose to V79 cells from extracellular 111In (i.e., from 111In located outside the target cell) was calculated to be 5.8 pGy/decay. This suggests that for an intracellular radiopharmaceutical, the radiation dose of consequence would be delivered by the low energy Auger electrons. In contrast, Auger electrons from an extracellular radiopharmaceutical could not directly damage the cell nucleus and therefore would not contribute to the radiation dose.     

Cell membrane is a more sensitive target than cytoplasm to dense ionization produced by auger electrons

To improve radioimmunotherapy with Auger electron emitters, we assessed whether the biological efficiency of (125)I varied according to its localization. A-431 and SK-OV-3 carcinoma cells were incubated with increasing activities (0-4 MBq/ml) of (125)I-labeled vectors targeting the cell membrane, the cytoplasm or the nucleus. We then measured cell survival by clonogenic assay and the mean radiation dose to the nucleus by assessing the cellular medical internal radiation dose (MIRD). The relationship between survival and the radiation dose delivered was investigated with a linear mixed regression model. For each cell line, we obtained dose-response curves for the three targets and the reference values (i.e., the dose leading to 75, 50 or 37% survival). When cell survival was expressed as a function of the total cumulative decays, nuclear (125)I disintegrations were more harmful than disintegrations in the cytoplasm or at the cell membrane. However, when survival was expressed as a function of the mean radiation dose to the nucleus, toxicity was significantly higher when (125)I was targeted to the cell membrane than to the cytoplasm. These findings indicate that the membrane is a more sensitive target than the cytoplasm for the dense ionization produced by Auger electrons. Moreover, cell membrane targeting is as cytotoxic as nuclear targeting in SK-OV-3 cells. We suggest that targeting the membrane rather than the cytoplasm may contribute to the development of more efficient radioimmunotherapies based on Auger electron radiation, also because most of the available vectors are directed against cell surface antigens.     

Relationship between radiation induced adaptive response in human fibroblasts and changes in chromatin conformation

Chromatin conformation changes in the normal human fibroblasts VH-10 were studied by the method of anomalous viscosity time dependence (AVTD). Gamma-irradiation of cells in a dose range of 0.1–3 Gy caused an increase in maximal viscosity of cell lysates. Conversely, irradiation of cells with low doses of 0.5 or 2 cGy resulted in a decrease in the AVTD peaks with a maximum effect approximately 40 min after irradiation. The same exposure conditions were used to study a possible adaptive effect of low doses, measured by changes in cell survival. A primary dose of 2 cGy caused significant modification of cell response to a challenge dose. Approximately 20% protection to challenge doses of 0.5 Gy (p < 0.003), 2 Gy (p < 0.02) and 2.5 Gy (p < 0.002) was observed. However, the direction of this effect (adaptation or synergism) was found to be dependent on a challenge dose. The combined effect of 2 cGy and 1 Gy was significantly synergistic, while no modification was observed for 1.5 Gy and 3 Gy. A partial correlation was found between the AVTD changes and cell survival when the combined effect of a primary dose of 2 cGy and challenge dose was examined. The dose of 2 cGy alone increased survival by 16% (p < 0.0003). These results suggest that the low-dose induced effects on survival may be related to chromatin reorganization.     

Feasibility study of the Fricke chemical dosimeter as an independent dosimetric system for the small animal radiation research platform (SARRP)

For the small animal radiation research platform (SARRP) with X-ray beams in the medium energy range (tube operating voltage at 220 kVp), reference dosimetry is based on the AAPM TG-61 recommendations following the in-phantom method. The objective of this study was to evaluate the feasibility of the Fricke solution as a dosimeter to determine the absorbed dose to water. Feasibility studies at this X-ray energy range are not widely available. We evaluated the accuracy, dose linearity and dose rate dependence in a comparison with an NE 2571 Farmer ionization chamber (IC) and measurements in water. The G(Fe³⁺) factor was calculated from the curve fitting of the chemical yields for two radioactive sources (¹⁹²Ir and ⁶⁰Co) and one X-ray system with a tube operating at 150 and 250 kVp. The same methodology was followed for the dependence of the G(Fe³⁺) value on the energy and the dose agreement assessment for 180 and 200 kVp in the SARRP. The Fricke system exhibits a good linear response over the range of 5–70 Gy and an accuracy better than 2% for a 2 Gy/min dose rate. The dose rate dependence is smaller than 1% for dose rates greater than 1 Gy/min. The dependence of the G(Fe³⁺) value on the energy is smaller than 0.41%, with dose agreements better than 2%. The feasibility of the dosimeter for measurements at high doses and high dose rates makes it a suitable tool for dosimetric verifications in several preclinical irradiation configurations.     

Kinetics of uptake, retention, and radiotoxicity of 125IUdR in mammalian cells: implications of localized energy deposition by Auger processes

The kinetics of uptake, retention, and radiotoxicity of 125IUdR have been studied in proliferating mammalian cells in culture. The radioactivity incorporated into the DNA is directly proportional to the duration of incubation and to the extracellular concentration of 125I. The rate of proliferation of cells is related to the intracellular radioactive concentration and is markedly reduced at medium concentrations greater than or equal to 0.1 mu Ci/ml. At 37% survival the high LET type cell survival curve is characterized by an uptake of 0.035 pCi/cell, and the cumulated mean lethal dose to the cell nucleus is about 80 rad compared to 580 rad of X-ray dose for this cell line. The strong cytocidal effects of the decay of 125I correlate with localized irradiation of the DNA by the low energy Auger electrons.     

Optimization of treatment planning for hypoxic tumours and re-modulation of radiation intensity in heavy-ion radiotherapy

AimThe purpose of this study is to optimize treatment planning in carbon ion radiotherapy, taking into account the effect of tumour hypoxia.BackgroundIn conventional hadron therapy, the goal is to create a homogenous dose in the tumour area and, thus, achieve a uniform cell survival level. Since the induction of a specific damage to cells is directly influenced by the level of hypoxia in the tissue, the varying oxygen pressure in the different regions of hypoxic tumours would disrupt the uniformity of the cell survival level.Materials and methodsUsing the Geant4 Monte Carlo Code, the physical dose profile and dose-averaged linear energy transfer were calculated in the tumour. Then, the oxygen enhancement ratio in different areas of the tumour were compared with different pressures.ResultsModulations of radiation intensities as well as energies of ion beams were calculated, both considering and disregarding the effect of hypoxia, and the required dose profiles were compared with each other. Cell survival levels were also compared between the two methods. An equation was obtained for re-modulating the beams in the presence of hypoxia, and radiation weighting factors were extracted for the beam intensities.ConclusionThe results show that taking the effect of hypoxia into account would cause the reduction of average doses delivered to the tumour tissues up to 1.54 times. In this regard, the required dose is reduced by 1.63 times in the healthy tissues before the tumour. This will result in an effective protection of healthy tissues around the tumour.     

Influence of 6 MV and 20 MV X-radiation dose rate on in vitro survive of the K-562 cell line

BackgroundAnalysis of the survival rate of cells after irradiation with a specified dose of X-radiation might be one of the basic foundations for assessment of biological implications of ionizing radiation. Investigation of the influence of X-radiation dose rate on cells was carried out in vitro using the SF2 test.AimThe aim of this study was to investigate the influence of X-radiation dose rate on the surviving fraction of the K-562 cell line for two photon energies of 6 MV and 20 MV.Materials/MethodsTo measure the cells' reaction to X-radiation of variable dose rate human leukaemic K-562 cells were used. In order to fulfil the main aim of the study, the cell line was subjected to irradiation at two different dose rates. Total dose applied at once was 2 Gy. A quantitative evaluation of cell survival rate was carried out at every step of the experiment using a clonogenic assay.ResultsHigh dose rate at the energy of 6 MV decreased the percentage of surviving cells to 23%, while lower dose rate decreased it only to 36%. A similar effect is observed at the energy of 20MV-namely at the higher dose rate the percentage of surviving cells is 18%, whereas at the lower one it is only 34%.ConclusionsThe experiment has shown that when using a lower dose rate, the biological effect of ionizing radiation is less pronounced. However, at a higher dose rate higher radiosensitivity of cells is observed.     

The effect of gamma-irradiation on the temperature dependence of D.C. electrical conductivity of dry bone.

The effect of gamma-irradiation with doses from 10 to 500 kGy on the electrical conductivity (g) of dry bone was studied. Temperature measurement of electrical conductivity were made from 393 to 533 K. The dependence obtained indicates the increase in g with temperature. An increase in irradiation dose resulted in a decreased g value for each dose up to temperature 462 K. Temperature 462 K was interpreted as the temperature of collagen melting point in dry bone. Above 462 K, g values were dose independent. A dose of 500 kGy shifted the melting point to lower temperature. In addition, the activation energy for the charge conduction process was calculated. Obtained values for electrical conductivity and activation energy were typical for dielectrics and indicated degradation of the organic component of bone.