EPR dosimetry with tooth enamel: A review

When tooth enamel is exposed to ionizing radiation, radicals are formed, which can be detected using electron paramagnetic resonance (EPR) techniques. EPR dosimetry using tooth enamel is based on the (presumed) correlation between the intensity or amplitude of some of the radiation-induced signals with the dose absorbed in the enamel. In the present paper a critical review is given of this widely applied dosimetric method. The first part of the paper is fairly fundamental and deals with the main properties of tooth enamel and some of its model systems (e.g., synthetic apatites). Considerable attention is also paid to the numerous radiation-induced and native EPR signals and the radicals responsible for them. The relevant methods for EPR detection, identification and spectrum analyzing are reviewed from a general point of view. Finally, the needs for solid-state modelling and studies of the linearity of the dose response are investigated. The second part is devoted to the practical implementation of EPR dosimetry using enamel. It concerns specific problems of preparation of samples, their irradiation and spectrum acquisition. It also describes how the dosimetric signal intensity and dose can be retrieved from the EPR spectra. Special attention is paid to the energy dependence of the EPR response and to sources of uncertainties. Results of and problems encountered in international intercomparisons and epidemiological studies are also dealt with. In the final section the future of EPR dosimetry with tooth enamel is analyzed.     

用电子顺磁共振波谱拟合方法估算牙釉质辐射剂量

目的 探讨运用电子顺磁共振(electron paramagnetic resonance, EPR)波谱拟合技术估算牙釉质EPR辐射剂量的准确性。方法 编制多成分叠加型EPR粉末波谱拟合软件,分别拟合牙齿本底信号(background signal,BS)和辐照诱发信号(radiation-induced signal,RS)的EPR波谱模型,用波谱模型叠加计算方法拟合出实际辐照后牙釉质的EPR波谱,从复合谱中提取出RS成分并计算其相对强度,建立剂量响应曲线,估算样品剂量,并将剂量估算结果与传统的波谱强度测量方法进行比较。结果 拟合获得的BS信号为单峰高斯线形粉末谱,g=2.0035,线宽Hpp=0.650-1.100 mT; RS信号为轴对称多晶粉末谱线形,其g_⊥=2.0018,g_‖＝1.9965,线宽Hpp=0.335-0.400 mT;分离BS与RS后得到的RS相对强度与辐照剂量呈线性相关,剂量响应方程为:y=240.74x+76 724(R²=0.9947),剂量估算结果相对误差期望值为0.13。结论 EPR波谱拟合方法在一定程度上提高了牙釉质辐射剂量估计的准确性和可信度。     

Metabolism in tooth enamel and reliability of retrospective dosimetry.

Most of the radiation centers in tooth enamel and bones are formed by the reactions COOH + gamma-->CO2- + H+, and CO2 + e- -->CO2-. These centers can be easily changed by metabolism and have a low stability in living organisms. The efficiency of CO2- radical production by irradiation is substantially different for teeth and bones in living rat, dead rat, and teeth and bone removed from rat. It is shown that the suitability of tooth enamel for retrospective dosimetry can be evaluated from the anisotropy of its EPR signals.     

EPR spectrum deconvolution and dose assessment of fossil tooth enamel using maximum likelihood common factor analysis.

In order to determine the components which give rise to the EPR spectrum around g = 2 we have applied Maximum Likelihood Common Factor Analysis (MLCFA) on the EPR spectra of enamel sample 1126 which has previously been analysed by continuous wave and pulsed EPR as well as EPR microscopy. MLCFA yielded agreeing results on three sets of X-band spectra and the following components were identified: an orthorhombic component attributed to CO2-, an axial component (CO3(3-)), as well as four isotropic components, three of which could be attributed to SO2-, a tumbling CO2- and a central line of a dimethyl radical. The X-band results were confirmed by analysis of Q-band spectra where three additional isotropic lines were found, however, these three components could not be attributed to known radicals. The orthorhombic component was used to establish dose response curves for the assessment of the past radiation dose, D(E). The results appear to be more reliable than those based on conventional peak-to-peak EPR intensity measurements or simple Gaussian deconvolution methods.     

Mechanically induced EPR signals in tooth enamel.

Sample preparation of tooth enamel for electron paramagnetic resonance (EPR) dosimetry usually involves mechanical operations. The present study shows that mechanical operations performed without water cooling generate a paramagnetic center inducing a stable isotropic EPR signal with g-value of 2.00320 and linewidth of about 0.1 mT. Using EPR spectrum simulation, the similarity between the mechanically induced signal and the signal generated when the enamel is heated in air at a temperature above 600 degrees C was investigated. Results indicate that the mechanically induced signal is related to sample temperature increase during mechanical friction.     

Gamma dose response of synthetic A-type carbonated apatite in comparison with the response of tooth enamel.

Synthetic A-type carbonated apatite samples were irradiated at room temperature with 60Co gamma rays. Their ESR spectra consist of the lines of CO2- and CO3- radicals of orthorhombic and axial symmetry. The measurements carried out immediately after sample irradiation showed that CO2- species are produced by decomposition of CO3- radicals. Intensity of the CO2- lines in the synthetic and enamel samples increases during the first 400 and 200 h after irradiation, respectively. The dependence of the EPR signal on the dose varies with carbonate content of the sample. The dose response curve for tooth enamel is steeper for the synthetic material.     

13C hyperfine interactions of CO2- in irradiated tooth enamel as studied by EPR.

The EPR spectrum of tooth enamel caused by ¹³C hyperfine interactions of the CO₂⁻ radical were studied on γ-irradiated powdered samples annealed for 40 min at different temperatures up to 250°C. The lineshape and hyperfine splitting of the spectra were found to depend on the annealing temperature. Experimental spectra were compared with calculated ones assuming that EPR spectra are formed by two CO₂⁻ species—axial (rotating) and orthorhombic (braked) radicals. We assumed that the axial CO₂⁻ radicals are centers located in perfect areas of the hydroxyapatite crystals of tooth enamel whereas the orthorhombic CO₂⁻ radicals are rotating centers which are braked by defects. The thermal treatment of enamel samples leads to defective annealing and transformation of the orthorhombic centers into axial ones. This results in an increasing axial CO₂⁻ radical contribution to the EPR spectrum with increase of annealing temperature.     

C hyperfine interactions of CO2 in irradiated tooth enamel as studied by EPR

The EPR spectrum of tooth enamel caused by ¹³C hyperfine interactions of the CO₂⁻ radical were studied on γ-irradiated powdered samples annealed for 40 min at different temperatures up to 250°C. The lineshape and hyperfine splitting of the spectra were found to depend on the annealing temperature. Experimental spectra were compared with calculated ones assuming that EPR spectra are formed by two CO₂⁻ species—axial (rotating) and orthorhombic (braked) radicals. We assumed that the axial CO₂⁻ radicals are centers located in perfect areas of the hydroxyapatite crystals of tooth enamel whereas the orthorhombic CO₂⁻ radicals are rotating centers which are braked by defects. The thermal treatment of enamel samples leads to defective annealing and transformation of the orthorhombic centers into axial ones. This results in an increasing axial CO₂⁻ radical contribution to the EPR spectrum with increase of annealing temperature.     

Elimination of the background signal in tooth enamel samples for EPR-dosimetry by means of physical-chemical treatment.

A method of elimination of the background EPR signal in tooth enamel is proposed. This method implies treatment of enamel powder by highly active reduction reagent hydrazine with subsequent washing out by ethanol-water solution. Such treatment results in reducing both the native background signal (which is assumed to be originated by the organic component) and the mechanical induced EPR signal in enamel. Testing of the efficiency of hydrazine treatment is made for different sizes of enamel powder. It is shown that the optimal results are obtained for a powder fraction of about 100-200 microm. The radiation-induced EPR signal in enamel is practically not changed after treatment by hydrazine.     

对辐照牙釉质EPR谱数学模拟的研究

目的　建立辐照牙釉质EPR谱数学模拟方法。方法　采用高斯函数一阶微商作为基本的模型函数 ,编写基于Marquardt Levenberg非线性最小二乘法曲线拟合算法的计算程序模拟辐照牙釉质EPR谱 ,并检验拟合精度。结果　对 2 70和 84 0mGy辐照的牙釉质样品的EPR复合谱 ,拟合后的残谱分别为 - 1 6 1± 2 3 5 9和 - 3 77± 2 4 94 ,残谱均值和标准差分别占各自峰高的 0 3%、3 8%和 0 4 %、3%。结论　这种算法和模型函数能很好地模拟辐照牙釉质EPR复合谱。     

Application of EPR retrospective dosimetry for large-scale accidental situation.

Above 3000 tooth enamel samples, collected at population of radioactive contaminated territories after Chernobyl accident, the Chernobyl liquidators, the retired military of high radiation risk and the population of control radiation free territories were investigated by EPR spectroscopy method in order to obtain accumulated individual exposure doses. Results of EPR spectra measurements are stored in data bank; enamel samples are also stored in order to provide the possibility to repeat the measurements in future. Statistical analysis of results has allowed to detect the contribution into EPR signal in tooth enamel due to the action of the natural background radiation, and the radioactive contamination of territory. In general, the average doses of external exposure of the population obtained with EPR spectroscopy of teeth enamel are consistent with results based on other methods of direct and retrospective dosimetry. Essential exceeding of the individual doses above the average level within the population groups was observed for some persons. That gave the possibility to detect the individuals with overexposure, which were included into groups for medical monitoring.     

Tooth enamel EPR dosimetry: sources of errors and their correction.

Some of the most important sources of systematic errors in dose determination using tooth enamel EPR spectroscopy and ways of reducing those errors are discussed. Enamel from the outside of the front teeth should not be used for dose determination because of induction of paramagnetic centers by solar light. The accuracy of the method in the low dose range is limited by variation in the shape of the EPR signal of unirradiated enamel, which can be described by an initial intrinsic signal and which varies for different samples with standard deviation of 20-30 mGy. The energy dependence of enamel sensitivity should be taken into account in the form of a correction factor. The value of this factor is estimated at 1.1-1.3 for real radiation fields in radiation contaminated territories. Variation in enamel sensitivity for different samples is shown to be within limits of 10-15% of the average value.     

Optimal registration conditions for tooth EPR dosimetry at low accumulated dose

The spectrum registration under rapid passage conditions (the second harmonic phase quadrature of the absorption signal) allows one to enhance substantially the sensitivity of tooth enamel and bone EPR dosimetry at a low accumulated dose. In the present work the dependencies of the radiation and background signals on EPR spectrometer parameters are described and the optimal conditions in RPM for EPR dosimetry are obtained.     

Parameters affecting EPR dose reconstruction in teeth.

The aim of this paper is to analyze the lower limit of detection (LLD), linearity of dose response, variation of radiation sensitivity between different tooth enamel samples, and time/temperature stability of EPR biodosimetry in tooth enamel. The theoretical LLD is shown to be 0.46 mGy, which is far lower than the measured value of about 30 mGy. The main issues to lowering LLD are the differentiation of the radiation-induced component against the total EPR spectrum and the complex nature of the dose dependence of the EPR signal. The following questions are also discussed in detail: need for exfoliated or extracted teeth from persons of interest, accounting for background radiation contribution; conversion of tooth enamel absorbed dose to effective dose; accounting for internal exposure specifically from bone-seeking radionuclides. Conclusions on future development of EPR retrospective biodosimetry are made.     

Temperature stability of carbonate groups in tooth enamel.

The tooth enamel EPR signal at g = 1.9973 consists of several components, which correspond to different functions of the spatial orientation of the CO2- radicals, have different temperature stability and different saturation curves. Using a new technique described in this paper, we have detected and investigated two groups of CO2- radicals with different thermal stability and saturation characteristics.     

Biophysical dose measurement using electron paramagnetic resonance in rodent teeth.

Electron paramagnetic resonance (EPR) dosimetry of human tooth enamel has been widely used in measuring radiation doses in various scenarios. However, there are situations that do not involve a human victim (e.g. tests for suspected environmental overexposures, measurements of doses to experimental animals in radiation biology research, or chronology of archaeological deposits). For such cases we have developed an EPR dosimetry technique making use of enamel of teeth extracted from mice. Tooth enamel from both previously irradiated and unirradiated mice was extracted and cleaned by processing in supersaturated KOH aqueous solution. Teeth from mice with no previous irradiation history exhibited a linear EPR response to the dose in the range from 0.8 to 5.5 Gy. The EPR dose reconstruction for a preliminarily irradiated batch resulted in the radiation dose of (1.4+/-0.2) Gy, which was in a good agreement with the estimated exposure of the teeth. The sensitivity of the EPR response of mouse enamel to gamma radiation was found to be half of that of human tooth enamel. The dosimetric EPR signal of mouse enamel is stable up at least to 42 days after exposure to radiation. Dose reconstruction was only possible with the enamel extracted from molars and premolars and could not be performed with incisors. Electron micrographs showed structural variations in the incisor enamel, possibly explaining the large interfering signal in the non-molar teeth.     

Retrospective radiation dosimetry using electron paramagnetic resonance in canine dental enamel.

Electron paramagnetic resonance (EPR) biodosimetry of human tooth enamel has been widely used for measuring radiation doses in various scenarios. We have now developed EPR dosimetry in tooth enamel extracted from canines. Molars and incisors from canines were cleaned by processing in supersaturated aqueous potassium hydroxide solution. The dosimetric signal in canine tooth enamel was found to increase linearly as a function of laboratory added dose from 0.44+/-0.02 to 4.42+/-0.22 Gy. The gamma radiation sensitivity of the canine molar enamel was found to be comparable to that of human tooth enamel. The dosimetric signal in canine enamel has been found to be stable up to at least 6 weeks after in vitro irradiation. A dosimetric signal variation of 10-25% was observed for canines ranging from in age 3 years to 16 year old.     

Changes in the paramagnetic centres in irradiated and heated dental enamel studied using electron paramagnetic resonance.

The EPR signals in dental enamel produced by radiation and by heat were studied. The inherent background signal at g = 2.005, and a radiation-produced signal at g = 2.002 have different saturation behaviour with microwave power, and this affords a method of signal optimization. Heating enamel at temperatures from 100 degrees C to 450 degrees C produces a range of radical species from g = 2.002 to g = 2.005, which have been characterized by their g-values, line widths and saturation behaviour. Standard dental drilling produces a range of radicals which appear to be similar to those produced by heat.     

Location of anthralin radical generation in mouse skin by UV-A irradiation: an estimation using microscopic EPR spectral-spatial imaging.

In vivo location of the anthralin radical generated in mouse skin by ultraviolet A (UV-A) irradiation was estimated by microscopic electron paramagnetic resonance (EPR) spectral-spatial imaging. An X-band EPR spectrometer equipped with specially designed high-power imaging coils and a TE-mode cavity was employed. The maximum field gradient used in this study was 6.77 mT/mm. Anthralin was applied to the dorsal skin of live mice, which were then exposed to UV-A irradiation. A broad singlet EPR spectrum (peak-to-peak line width = 0.6 mT and g = 2.004) was obtained. Microscopic EPR spectral-spatial imaging of the skin tissue showed that the anthralin radical was located mainly in the epidermis (27 microm from the skin surface). This result was consistent with the finding that the proportions of the radical in the dermis and epidermis were about 15% and 85%, respectively.     

Comparison of sample preparation and signal evaluation methods for EPR analysis of tooth enamel.

In dose reconstruction by EPR dosimetry with teeth various methods are applied to prepare tooth enamel samples and to evaluate the dosimetric signal. A comparison of seven frequently used methods in EPR dosimetry with tooth enamel was performed. The participating Institutes have applied their own procedure to prepare tooth enamel samples and to evaluate the dosimetric signal. The precision of the EPR measurement and the dependence of the estimated dosimetric signal with irradiation up to 1000 mGy were compared. The obtained results are consistent among the different methods. The reproducibility of the dosimetric signal and its estimated relation with the absorbed dose was found to be very close for the applied methods with one possible exception.