2.1. Design and configuration of the wearable resonator
A wearable resonator on a tooth was designed for in-vivo EPR spectroscopy in conjunction with a microwave circuit. Figure 1 shows the conceptual design of the wearable resonator applied to a human head phantom. The wearable resonator consists of three components: an attachable surface coil, a flexible transmission line, and a coupling circuit. Each component is tailored to meet specific requirements for effective in-vivo tooth dosimetry. The attachable surface coil serves the crucial function of generating a magnetic field to excite an EPR signal within the subject. It is designed to adhere securely to the tooth surface using adhesive, ensuring consistent positioning with the tooth despite potential movement by the subject. This flexibility is essential to maximize adhesion on the irregular surface of the tooth. Despite the potential movement of the attachable surface coil with the tooth, the opposite end of the coil’s transmission line is connected to the coupling circuit. This arrangement necessitates significant flexibility in the transmission line to accommodate varying positions. Additionally, the coupling circuit should feature a wide tunable frequency range to effectively compensate for impedance perturbation caused by subject movement. The proposed resonator is specifically designed with a resonant frequency of 1.15 GHz to align with the specifications of the RF bridge used in this study.
A circuit diagram of the wearable resonator is shown in Fig. 2. To facilitate the use of the feedback system, we adopted a modified version of a design reported previously [22]. The attachable surface coil, acting as a parallel inductance LP, was custom-made from a flexible printed circuit board (FPCB). The FPCB consisted of a substrate and a copper layer, etched into the form of a surface coil with a thickness of several tens of micrometers. A double-sided medical-grade adhesive (3M™ 1577 Double Coated Medical Tape) was employed to attach the FPCB-based surface coil to the tooth surface. This adhesive exhibits strong bonding strength while also being readily detachable without causing damage when moderate force is applied. To achieve parallel LC resonance with the surface coil and minimize dielectric loss, a copper-laminated dielectric substrate was cut into 3 mm × 3 mm pieces (Cuflon CF-A-3–7–7, Polyflon Company, Norwalk, CT, USA). A commercial flexible coaxial cable was cut to a length five times the quarter wavelength to serve as parallel transmission lines (225 mm), providing sufficient length to accommodate a wider range of subject movement (SLC-6FT-SMSM+, Mini-Circuits, Brooklyn, NY, USA). A reduction in the coaxial cable length, specifically to either one or three quarter-wavelengths, can result in decreased the transmitted signal attenuation. However, this modification concurrently constrains the permissible range of tooth displacement. The coupling circuit part consists of variable capacitors, CT and CM (PTIC, TCP-5082, ON Semiconductor, Phoenix, USA), connected in series and parallel, respectively. This section is connected to the EPR bridge as shown in Fig. 2. To apply bias voltages VT and VM for adjusting the variable capacitance, RC low-pass filters were used. An RF choke, implemented using a commercial inductor (100 nH, LQW2BASR10J00L, Murata, Kyoto, Japan), served as a low-pass filter to eliminate microwave frequencies under 1 GHz. The bias voltages VT and VM of the coupling circuit part were controlled by an automatic control circuit (ACC), maintaining the critical coupling state of the resonator to minimize reflected RF power [22]. All components were soldered using 3% silver-based solder. The coupling circuit part was shielded with a 6-mm-thick copper case to minimize the effect of the modulation magnetic field, which has a frequency of 21.6 kHz.
To evaluate the resonator characteristics, we used a network analyzer (E5061B, Keysight Technologies, USA) to measure the scattering matrix parameter S11. Measurements were recorded at 1601 data points over a span of 50 MHz. From these measurements, we estimated the resonance frequency and bandwidth of the resonator. The quality factor, which is proportional to the EPR sensitivity of the resonator, was calculated by dividing the resonance frequency by the bandwidth. Additionally, we quantified the shift in resonance frequency resulting from tooth movement.
2.2. Simulation of the attachable surface coil
The attachable surface coil LP and parallel capacitor CP, as shown in Fig. 2, are analyzed by finite element method in ANSYS HFSS. In simulation model, critical coupling of the resonator was achieved by fine-tuning CT and CM, which are the tuning and matching capacitances, respectively, within the range from 2 to 8 pF. Sinusoidal power was applied to the resonator through a 50 Ω impedance microwave port. The S11 was calculated using linear network analysis across a frequency sweep range of 0 to 3 GHz to estimate the resonance frequency of the resonator. Finally, the dimensions of the attachable surface coil LP and parallel capacitor CP were optimized to achieve a resonance frequency close to 1.15 GHz for the wearable resonator.
Figure 3 illustrates an incisor tooth with the attachable surface coil configured in ANSYS HFSS. The tooth model was obtained using a micro-CT image of an extracted central upper incisor (Skyscan 1275, Bruker SkyScan, Kontich, Belgium). Enamel, dentin, and pulp of the tooth were segmented based on the Hounsfield unit obtained from the micro-CT scan. The electrical properties of each material were based on characteristics reported in a previous study [33]. Out of 12 micro-CT-based 3D models of the upper central incisor, the simulation utilized the sample closest to the average size. The plateau region of the enamel surface area on the 3D incisor model measured 5.5 mm in width and 9.5 mm in height; these dimensions are suitable for coverage by the FPCB without significant deformation. To accommodate the curvature of the enamel surface, the FPCB was bent and adhered accordingly. For the design optimization of the FPCB, finite element analysis was conducted within feasible ranges of design parameters for FPCB fabrication. The inner diameter and trace width were varied from 4 to 6 mm and 0.4 to 2 mm, respectively, to maximize the efficiency of magnetic field generation in terms of filling factor (η). The filling factor, a crucial optimization metric for resonators discussed in previous literature on instrumental EPR [34], was calculated within the software based on the following definition:
$$\:\text{f}\text{i}\text{l}\text{l}\text{i}\text{n}\text{g}\:\text{f}\text{a}\text{c}\text{t}\text{o}\text{r},\:{\eta\:}=\frac{{\int\:}_{enamel}^{\:}{B}_{1,\perp\:}^{2}dV}{{\int\:}_{all\:objects}^{\:}{B}_{1}^{2}dV},$$
where \(\:{B}_{1}\) denotes the magnetic field generated by the resonator. \(\:{B}_{1,\perp\:}\) denotes the component of B1 orthogonal to an external static magnetic field B0 (horizontal axis in Fig. 3) and is crucial for EPR signal intensity. Essentially, the filling factor quantifies the ratio of the magnetic field concentrated within the enamel volume to the total magnetic field generated.
2.3. Ex-vivo EPR measurements of the wearable resonator
To evaluate the performance of the fabricated wearable resonator, experimental validation was conducted using an EPR spectrometer interfaced with an inhouse-developed EPR spectrometer [22, 35, 36]. The wearable resonator received tuning and matching voltages (VT and VM, respectively) through a previously developed ACC. A microwave power of 100 mW was applied from a signal generation in EPR bridge, with amplitude modulation of the magnetic field intensity set at 0.4 mT for field modulation. The EPR spectrum was obtained and averaged over 30 s, comprising 10 repetitions of a 3-s sweep. For tooth radiation dosimetry, a single extracted tooth sample irradiated with 50 Gy using 220 kVp X-ray, calibrated according to the dosimetric protocol of the American Association of Physicists in Medicine [37], was used. The study was approved by the Institutional Review Board (IRB) of Seoul National University Hospital (Approval No. 2206/027-1330). During EPR measurement, the surface coil of the wearable resonator was attached to the tooth enamel. The coil was positioned such that its axis coincided with the center of the tooth’s horizontal axis, and its inner side aligned with the edge of the tooth enamel, as shown in Fig. 3. For reference, a 1 mM 15N-substituted perdeuterated 2,2,6,6-tetramethyl-4-oxopi peridine-1-oxyl sample (15N-PDT, CDN Isotopes, Quebec, Canada) sealed in a Teflon tube was simultaneously attached to the surface coil for measurement. The acquired EPR spectrum was analyzed using a home-built EPR signal processing software [36]. The RIS intensity was evaluated based on peak-to-peak amplitude after non-linear fitting the EPR line shape. The RIS amplitudes of the wearable resonator and a conventional rigid resonator [22] were compared.
To assess the impact of tooth displacement, an experimental setup was employed to manipulate the resonator position. Previous work has identified that the tooth displacement along the central axis of the resonator coil, moving away from the tooth’s frontal surface, is the primary determinant of RIS amplitude [7]. In this investigation, the tooth’s position remained constant while the resonator was displaced along the central axis from 0 to 2 mm in intervals of 0.5 mm using a precision moving stage. At each displacement, five EPR spectra were measured using both the rigid and wearable resonators.
An additional experiment was performed to verify the uncertainty and reproducibility of the wearable resonator. To assess the positioning uncertainty of both the resonator and reference sample tube, repetitive measurements were conducted on the irradiated tooth. After acquiring five spectra, the surface coil and sample tube were detached from the tooth and repositioned for another measurement. A total of 30 independent EPR measurements were conducted, and in each measurement, five EPR spectra were acquired for quantitative analysis.
2.4. In-vivo EPR measurements of the wearable resonator
To validate the developed wearable resonator in an in-vivo setting, a single volunteer, who is not irradiated by ionizing radiation, underwent EPR measurements. The EPR spectrum was acquired with and without the ACC. The SNR ratio was determined by dividing the signal amplitude by twice the standard deviation of the noise level. The study was approved by the IRB of Seoul National University (Approval No. 2207/002–002). Figure 4 illustrates the procedural steps for applying the wearable resonator to the subject’s tooth. Initially, the subject’s chin was placed on a chin rest integrated into the magnet system, and the subject’s head was secured using a head restraint to ensure immobilization. The attachable surface coil of the wearable resonator was then affixed to the surface of the subject’s upper central incisor. To identify RIS center in EPR spectrum, a reference sample tube should be measured simultaneously. Before analyzing the EPR spectrum, the reference sample tube was placed over the surface coil on the subject’s tooth and wrapped around the upper dentition with surgical tape, as shown in Fig. 4 (d).