Biological characteristics of carbon-ion therapy

Purpose: Radiotherapy using charged and/or high-linear energy transfer (LET) particles has a long history, starting with proton beams up to now carbon-ions. Radiation quality of particle beams is different from conventional photons, and therefore the biological effects of high-LET irradiation have attracted scientific interests of many scientists in basic and clinical fields. A brief history of particle radiotherapy in the past half-century is followed by the reviewed biological effectiveness of high-LET charged particles.

Results: The latter includes 54 papers presenting 506 RBE (relative biological effectiveness) values for carbon ions and a total of 290 RBE values for other ions identified from 48 papers. By setting a selection window of LET up to 100 keV/μm, we fitted a linear regression line to an LET-RBE relation. The resulting slope of the regression line had a dimension of μm/keV, and showed different steepness for different cells/tissues and endpoints as well. The steepest regression was found for chromosome aberration of human malignant melanoma while the shallowest was for apoptosis of rodent cells/tissue. Both tumour and normal tissue showed relatively shallower slopes than colony formation.

Conclusions: In general, there is a large variation of slope values, but the majority (25 out of 29 values) of data was smaller than 0.05 μm/keV.     

Biological effectiveness of 12C and 20Ne ions with very high LET

Purpose: To determine the relationship between the relative biological effectiveness (RBE) for cell inactivation and linear energy transfer (LET) in the Bragg peak region of ¹²C and ²⁰Ne ions.

Materials and methods: Chinese hamster ovary (CHO-K1) cells were exposed to high LET ¹²C (33.2 MeV, 20.3 MeV, 9.1 MeV at cell entrance) and ²⁰Ne ions (56.2 MeV, 34.7 MeV, 15 MeV at cell entrance) and to low LET x-rays. Technical details of the irradiation facility are presented which is based on the Monte Carlo simulation of the lateral spread of heavy ions as a result of the multiscattering small-angle process in physical conditions of the experimental set-up.

Results: RBE has been measured for LET values close to the Bragg peak maximum, i.e., 440–830 keV/μm for ¹²C and for 1020–1600 keV/μm for ²⁰Ne ions. RBE values at several levels of survival were estimated and were found to decrease with increasing LET. The inactivation cross sections were calculated from the final slope of dose-response curves and were found to increase with increasing LET.

Conclusions: The RBE decreases with increasing LET in the range between 440 and 1600 keV/μm for the two types of radiations forming a single line when plotted together, pointing towards LET as the single determinant of RBE. The inactivation cross section describing the killing efficiency of a single particle at the end of particle range comes close to the size of the cell nucleus.     

A small fish model for quantitative analysis of radiation effects using visualized thymus responses in GFP transgenic medaka

Abstract

Purpose: The aim of this study was to establish a new method of real-time, in vivo detection of radiation damage and recovery.

Methods: The thymus was observed under fluorescent light in a green fluorescent protein transgenic medaka. After irradiation, medaka thymus images were analyzed to quantify the effects of radiation by measuring changes in thymus size. A single acute irradiation of X-rays (0–30?Gy) or heavy Fe ions (0–10?Gy) was delivered to the medaka. Images were captured 0, 1, 2, 3, 5, 7, 11, and 21 d after irradiation. Dose-response assessment was conducted to provide a direct measurement of the effects of the radiation.

Conclusion: A biomonitoring system to detect the effects of radiation in real time was established. Using this system, the threshold doses for the induction of thymic atrophy by acute X-rays and Fe ions were 2–5?Gy and 0.5–1?Gy, respectively. The Relative Biological Effectiveness (RBE) of Fe-ion to X-rays was estimated to be around 3. This system may be used to evaluate the risk from concurrent exposure to hazards, such as chemicals and radiation, and for aging research.     

Dose-response and relative biological effectiveness of fast neutrons: Induction of apoptosis and inhibition of neurogenesis in the hippocampus of adult mice

Purpose:?Our study compared the effects of high linear energy transfer (LET) fast neutrons on the induction of apoptosis and reduction of neurogenesis in the hippocampus of adult ICR mice with those of low-LET ⁶⁰Co γ-rays, to evaluate the relative biological effectiveness (RBE) of fast neutrons in the adult hippocampal dentate gyrus (DG).

Materials and Methods:?The mice were exposed to 35 MeV fast neutrons or ⁶⁰Co γ-rays. We evaluated acutely the incidence of apoptosis and expression of Ki-67 (a protein marker for cell proliferation originally defined by the monoclonal antibody Kiel-67) and doublecortin (DCX: an immature progenitor neuron marker) in the hippocampus after a single whole-body irradiation.

Results:?The number of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labelling (TUNEL)-positive apoptotic nuclei in the DG increased and both Ki-67- and DCX-positive cells declined in a dose-dependent pattern, with fast neutrons or γ-rays. In the hippocampus, which showed an apoptosis frequency between 2 and 8 per DG, the RBE of fast neutrons was approximately 1.9. Additionally, the inhibitory effects of fast neutrons on the expression frequencies of Ki-67 (4–8) and DCX (8–32) were approximately 3.2 and 2.5 times, respectively, the effects of γ-rays at the same dose.

Conclusions:?Increased apoptotic cell death and decreased neurogenesis in the hippocampal DG were seen in a dose-dependent pattern after exposure to fast neutrons and γ-rays. In addition, the different rate of hippocampal neurogenesis between different radiation qualities may be an index of RBE.     

Monte Carlo simulations of therapeutic proton beams for relative biological effectiveness of double-strand break

Abstract

Purpose: The relative biological effectiveness (RBE) values relative to ⁶⁰Co for the induction of double-strand breaks (DSB) were calculated for therapeutic proton beams. RBE-weighted absorbed doses were determined at different depths in a water phantom for proton beams.

Materials and methods: The depth-dose distributions and the fluence spectra for primary protons and secondary particles were calculated using the FLUKA (FLUktuierende KAskade) MC (Monte Carlo) transport code. These spectra were combined with the MCDS (Monte Carlo damage simulation) code to simulate the spectrum-averaged yields of clustered DNA lesions. RBE for the induction of DSB were then determined at different depths in a water phantom for the unmodulated and modulated proton beams.

Results: The maximum RBE for the induction of DSB at 1 Gy absorbed dose was found about 1.5 at 0.5 cm distal to the Bragg peak maximum for an unmodulated 160 MeV proton beam. The RBE-weighted absorbed dose extended the biologically effective range of the proton beam by 1.9 mm. The corresponding maximum RBE value was inversely proportional to the proton beam energy, reaching a value of about 1.9 for 70 MeV proton beam. For a modulated 160 MeV proton beam, the RBE weightings were more pronounced near the spread-out Bragg peak (SOBP) distal edge.

Conclusions: It was demonstrated that a fast MCDS code could be used to simulate the DNA damage yield for therapeutic proton beams. Simulated RBE for the induction of DSB were comparable to RBE measured in vitro and in vivo. Depth dependent RBE values in the SOBP region might have to be considered in certain treatment situations.     

低剂量氚相对生物效能的实验研究与建议

笔者从辐射防护的角度概述了核聚变燃料氚低剂量照射下的相对生物效能 (RBE）的研究。选择指数递减剂量率和恒定剂量率2种氚照射方式,观察研究以下生物学指标：卵母细胞和精母细胞的显性致死突变率,显性骨骼突变率,初级卵母细胞和精原细胞的存活率,以及外周血淋巴细胞和胎肝嗜多染红细胞微核细胞率。计算2种氚照射方式下的RBE值,并分析RBE的影响因素。结果显示,在累积剂量为0.2、0.3、0.4、0.5、0.6 Gy/10 d的条件下,指数递减剂量率和恒定剂量率2种氚照射方式下的RBE值为2.9~4.2。为了辐射防护的目的,建议将低传能线密度（LET）辐射对生物群的RBE值设定为3.0~3.5。如果估计暴露于氚β粒子或其他低LET辐射或接近导出考虑参考水平下,则可能需要采用较高的RBE值进行评估,以更为准确地估计辐射的危险度。     

Tumor induction in mice after local irradiation with single doses of either carbon-ion beams or gamma rays

Abstract

Purpose: To determine the dose-dependent relative biological effectiveness (RBE) for tumor prevalence in mice receiving single localized doses to their right leg of either carbon ions (15, 45 or 75 keV/μm) or 137Cs gamma rays.

Methods and materials: A total of 1647 female C3H mice were irradiated to their hind legs with a localized dose of either reference gamma rays or 15, 45 or 75 keV/μm carbon-ion beams. Irradiated mice were evaluated for tumors twice a month during their three-year life span, and the dimensions of any tumors found were measured with a caliper. The tumor induction frequency was calculated by Kaplan-Meier analysis.

Results: The incidence of tumors from 50 Gy of 45 keV/μm carbon ions was marginally higher than those from 50 Gy of gamma rays. However, 60 Gy of 15 keV/μm carbon ions induced significantly fewer tumors than did gamma rays. RBE values of 0.87 + 0.12, 1.29 + 0.08 or 2.06 + 0.39 for lifetime tumorigenesis were calculated for 15, 45 or 75 keV/μm carbon-ion beams, respectively. Fibrosarcoma predominated, with no Linear Energy Transfer (LET)-dependent differences in the tumor histology. Experiments measuring the late effect of leg skin shrinkage suggested that the carcinogenic damage of 15 keV/μm carbon ions would be less than that of gamma rays.

Conclusions: We conclude that patients receiving radiation doses to their normal tissues would face less risk of secondary tumor induction by carbon ions of intermediate LET values compared to equivalent doses of photons.     

重离子生物效应的研究进展

重离子是一种具有特殊物理性质的带电离子,在射程末端可形成Bragg峰。与X射线、γ射线等低传能线密度（LET）射线相比具有较高的相对生物效应。笔者从细胞水平和分子水平简要介绍了重离子的辐射生物效应,以及与低LET射线的主要区别。     

Neurocytotoxic effects of iron-ions on the developing brain measured in vivo using medaka (Oryzias latipes), a vertebrate model

Abstract

Purpose: Exposure to heavy-ion radiation is considered a critical health risk on long-term space missions. The developing central nervous system (CNS) is a highly radiosensitive tissue; however, the biological effects of heavy-ion radiation, which are greater than those of low-linear energy transfer (LET) radiation, are not well studied, especially in vivo in intact organisms. Here, we examined the effects of iron-ions on the developing CNS using vertebrate organism, fish embryos of medaka (Oryzias latipes).

Materials and methods: Medaka embryos at developmental stage 28 were irradiated with iron-ions at various doses of 0–1.5 Gy. At 24 h after irradiation, radiation-induced apoptosis was examined using an acridine orange (AO) assay and histologically. To estimate the relative biological effectiveness (RBE), we quantified only characteristic AO-stained rosette-shaped apoptosis in the developing optic tectum (OT). At the time of hatching, morphological abnormalities in the irradiated brain were examined histologically.

Results: The dose–response curve utilizing an apoptotic index for the iron-ion irradiated embryos was much steeper than that for X-ray irradiated embryos, with RBE values of 3.7–4.2. Histological examinations of irradiated medaka brain at 24 h after irradiation showed AO-positive rosette-shaped clusters as aggregates of condensed nuclei, exhibiting a circular hole, mainly in the marginal area of the OT and in the retina. However, all of the irradiated embryos hatched normally without apparent histological abnormalities in their brains.

Conclusion: Our present study indicates that the medaka embryo is a useful model for evaluating neurocytotoxic effects on the developing CNS induced by exposure to heavy iron-ions relevant to the aerospace radiation environment.     

Cell cycle arrest and aberration yield in normal human fibroblasts. II: Effects of 11 MeV u−1 C ions and 9.9 MeV u−1 Ni ions

Purpose: To investigate further the relationship between high linear energy transfer (LET) induced cell cycle arrests and the yield of chromosome aberrations observable in normal human fibroblasts at the first post-irradiation mitosis.

Materials and methods: Normal human fibroblasts (AG01522C) were exposed in G0/G1 to either 11 MeV u^?1 C ions (LET = 153.5 keV μm^?1) or 9.9 MeV u^?1 Ni ions (LET = 2455 keV μm^?1), subcultured in medium containing 5-Bromo-2'-deoxyuridine (BrdU) and at multiple time-points post-irradiation the yield of chromosomal damage, the mitotic index and the cumulative BrdU-labelling index were determined. Furthermore, a mathematical approach was used to analyse the entire cell population.

Results: Following high LET exposure normal fibroblasts suffer a transient delay into S-phase and into mitosis as well as a prolonged, probably permanent cell cycle arrest in the initial G0/G1-phase. Cells that reach the first mitosis at early times carried less aberrations than those collected at later times indicating a relationship between cell cycle delay and the number of aberrations. However, with respect to the whole cell population, only a few aberrant fibroblasts are able to progress to the first mitosis. For all endpoints studied the relative biological effectiveness (RBE) of C ions is in the range of 2 – 4, while for Ni ions RBE < 1 is estimated. In contrast, when compared on a per particle basis Ni ions with the higher ionization density were found to be more effective.

Conclusions: Detailed analysis of the data demonstrates that the number of fibroblasts at risk for neoplastic transformation is significantly reduced by a chronic cell cycle arrest in the initial G0/G1-phase and, for the first time, the LET-dependence of this effect has been shown.     

Interaction between the biological effects of high- and low-LET radiation dose components in a mixed field exposure

Abstract

Purpose: The relative biological effectiveness of two epithermal neutron sources, a reactor based source at Studsvik, Sweden, and a proton accelerator-based source in Birmingham, UK, was studied in relation to the proportional absorbed dose distribution as a function of neutron energy. Evidence for any interactions between the effects of biological damage induced by high- and low-linear energy transfer (LET) dose components, in this ‘mixed field’ irradiation, was also examined

Materials and methods: Clonogenic survival in Chinese Hamster-derived V79 cells was used to assess biological effectiveness in this study. Cells were irradiated in suspension at 4°C at depths of 20, 35, 50 and 65 mm in a water phantom. This prevented the repair of sublethal damage, predominantly that produced by both incident and induced γ-rays in the field, over the variable periods of exposure required to irradiate cells with the same total absorbed dose. Cell survival, as a function of the absorbed radiation dose and depth in the phantom, was compared with Monte Carlo N-Particle (MCNP) calculations of the proportional absorbed dose distribution as a function of neutron energy for the two sources.

Results: In terms of the dose-related reduction in clonogenic cell survival, the epithermal neutron source at Studsvik was more biologically effective than the Birmingham source at all depths considered in the phantom. Although the contribution from the high-LET dose component was greater for the Studsvik source at 20 mm depth in the phantom, at greater depths the dose contribution from the high-LET dose component at Studsvik overlap with those for the Birmingham source. However, the most striking difference is in the fast neutron component to the dose of the two sources, neutron energies > 1 MeV were only associated with the Studsvik source. The relative biological effectiveness (RBE) of both sources declined slightly with depth in the phantom, as the total high-LET dose component declined. The maximum source RBE for Studsvik was 2.70 ± 0.50 at 20 mm; reduced to 2.10 ± 0.35 at depths of 50 and 65 mm. The corresponding values for Birmingham were 1.68 ± 0.25 and 1.31 ± 0.19, all values relate only to the surviving fraction of V79 cells at 37%, since RBE values are only applicable to the selected endpoint. Based on a dose reduction factor (DRF) of 1.0 for the total low-LET component to the absorbed dose, the RBE values for the high-LET dose component (fast neutrons and induced protons from the nitrogen capture reaction) was 14.5 and 7.05 for the Studsvik and Birmingham neutron sources, respectively. This is well outside the range of RBE historically reported values for V79 cells for the same level of cell survival for fast neutrons. The calculation of RBE values, based on the proportional absorbed dose distribution as a function of neutron energy, from historical data, and using a RBE of 1.8 for the dose from the nitrogen capture reaction, suggests RBE values for the total high-LET dose component of 3.1–2.8 and 2.5–2.0 for Studsvik and Birmingham, respectively, values again declining with depth in the phantom.

Conclusions: The overall biological effectiveness of the mixed field irradiation from an epithermal neutron sources depends on the composition and quality of the different dose components. The experimentally derived RBE values for the total high-LET dose components in these ‘mixed field’ irradiations are well in excess of historical data for fast neutrons. The difference between the historically expected and the observed RBE values is attributed to the interactions between the damage produced by high- and low-LET radiation.     

Doses and lung cancer risks from exposure to radon and plutonium

Abstract

Purpose: Epidemiological studies of the French uranium miners and the plutonium workers at the Mayak nuclear facility have provided excess relative risk (ERR) estimates per unit absorbed lung dose from alpha radiation. The aim of this paper was to review these two studies and to derive values of the relative biological effectiveness (RBE) of alpha particles for the induction of lung cancer.

Materials and methods: We examined and compared the dosimetry assumptions and methodology used in the epidemiological studies of uranium miners and the plutonium workers. Values of RBE were obtained by comparing risk coefficients including comparison of lifetime risks for a given population. To do this, preliminary calculations of lifetime risks following inhalation of plutonium were carried out.

Results and conclusions: Published values of risk per unit dose following inhalation of radon progeny and plutonium were in agreement despite the very different dose distributions within the lungs and the different ways the doses were calculated. Values of RBE around 10–20 were obtained by comparing ERR values, but with wide uncertainty ranges. Comparing lifetime risks gave similar values (10, 19 and 21). This supports the use of a radiation weighting factor of 20 for alpha particles for radiation protection purposes.     

Radiobiological evaluation and correlation with the local effect model (LEM) of carbon ion radiation therapy and temozolomide in glioblastoma cell lines

Purpose:?To investigate the cytotoxic effect of high linear-energy transfer (LET) carbon irradiation on glioblastoma cells lines in combination with temozolomide (TMZ).

Methods and materials:?The cell lines U87-MG expressing wild-type p53 and LN229 expressing both mutant and wild-type p53 were irradiated with monoenergetic carbon ion beams (LET 172 keV/μm) or an extended Bragg peak (LET 103 keV/μm) after treatment with 10 μM or 20 μM TMZ. Cytotoxicity was measured by a clonogenic survival assay, and cell growth as well as cell cycle progression, were examined.

Results:?The p53 mutant was more sensitive to X-ray irradiation than the p53 wild type cell line, which was also expressed in a shorter G2 block. High LET carbon ions show an increased biological effectiveness in both cell lines, which is consistent with the predictive calculations by the Local Effect Model (LEM) introduced by Scholz et al. The cell line LN229 was more sensitive to TMZ treatment than the U87MG cell line expressing wild-type p53 only. The combination of TMZ and irradiation showed an additive effect in both cell lines.

Conclusion:?High LET carbon ion irradiation is significantly more effective for glioblastoma cell lines compared to photon irradiation. An additional treatment with TMZ may offer a great chance especially for several tumor types.     

头颈部肿瘤三维适形放射治疗中的质量保证

目的探讨头颈部肿瘤3D—CRT实施过程的质量控制(QC)方法。方法采用拓能系统的固定装置、计划系统、激光灯、弓形尺、头部模型(简称头模)模拟3D—CRT的实施过程进行质量控制分组实验。结果复位标记标在固定装置以及面罩开窗处患者皮肤上的B组，实施过程误差小于复位标志标注在面罩上的A组。能达到3D—CRT过程QC要求。结论开展头颈部肿瘤3D—CRT必须合理使用头部固定装置。严格按本文规定标准方法操作可达到3D—CRT的实施过程的QC要求。