Optical super-resolution imaging by high-index microspheres embedded in elastomers.

We demonstrated feasibility of super-resolution imaging through high-index microspheres embedded in transparent elastomers. We performed imaging, with resolution improvement by a factor of two, by using implanted barium titanate glass microspheres (diameters ∼30-150 µm and refractive index ∼1.9-2.1) in a thin film of polydimethylsiloxane elastomer placed over the specimen. Microsphere-assisted imaging technique is a promising candidate for applications in cancer research. As a proof-of-principle, we used microsphere-assisted imaging technique for the observation of radiation-induced γ-H2AX foci formation in U87 human glioblastoma cells irradiated by clinical proton beams.

First experimental measurements of 2D microdosimetry maps in proton therapy.

BACKGROUND: Empirical data in proton therapy indicate that relative biological effectiveness (RBE) is not constant, and it is directly related to the linear energy transfer (LET). The experimental assessment of LET with high resolution would be a powerful tool for minimizing the LET hot spots in intensity-modulated proton therapy, RBE- or LET-guided evaluation and optimization to achieve biologically optimized proton plans, verifying the theoretical predictions of variable proton RBE models, and so on. This could impact clinical outcomes by reducing toxicities in organs at risk. PURPOSE: The present work shows the first 2D LET maps obtained at a proton therapy facility using the double scattering delivery mode in clinical conditions by means of new silicon 3D-cylindrical microdetectors. METHODS: The device consists of a matrix of 121 independent silicon-based detectors that have 3D-cylindrical electrodes of 25-µm diameter and 20-µm depth, resulting each one of them in a well-defined micrometric radiation sensitive volume etched inside the silicon. They have been specifically designed for a hadron therapy, improving the performance of current silicon-based microdosimeters. Microdosimetry spectra were obtained at different positions of the Bragg curve by using a water-equivalent phantom along an 89-MeV pristine proton beam generated in the Y1 proton passive scattering beamline of the Orsay Proton Therapy Centre (Institut Curie, France). RESULTS: Microdosimetry 2D-maps showing the variation of the lineal energy with depth in the three dimensions were obtained in situ during irradiation at clinical fluence rates (∼108  s-1  cm-2 ) for the first time with a spatial resolution of 200 µm, the highest achieved in the transverse plane so far. The experimental results were cross-checked with Monte Carlo simulations and a good agreement between the spectra shapes was found. The experimental frequency-mean lineal energy values in silicon were 1.858 ± 0.019 keV µm-1 at the entrance, 2.61 ± 0.03 keV µm-1 at the proximal distance, 4.97 ± 0.05 keV µm-1 close to the Bragg peak, and 8.6 ± 0.1 keV µm-1 at the distal edge. They are in good agreement with the expected trends in the literature in clinical proton beams. CONCLUSIONS: We present the first 2D microdosimetry maps obtained in situ during irradiation at clinical fluence rates in proton therapy. Our results show that the arrays of 3D-cylindrical microdetectors are a reliable microdosimeter to evaluate LET maps not only in the longitudinal axis of the beam, but also in the transverse plane allowing for LET characterization in three dimensions. This work is a proof of principle showing the capacity of our system to deliver LET 2D maps. This kind of experimental data is needed to validate variable proton RBE models and to optimize LET-guided plans.

Microdosimetry performance of the first multi-arrays of 3D-cylindrical microdetectors.

The present work reports on the microdosimetry measurements performed with the two first multi-arrays of microdosimeters with the highest radiation sensitive surface covered so far. The sensors are based on new silicon-based radiation detectors with a novel 3D cylindrical architecture. Each system consists of arrays of independent microdetectors covering 2 mm[Formula: see text]2 mm and 0.4 mm[Formula: see text]12 cm radiation sensitive areas, the sensor distributions are arranged in layouts of 11[Formula: see text]11 microdetectors and 3[Formula: see text]3 multi-arrays, respectively. We have performed proton irradiations at several energies to compare the microdosimetry performance of the two systems, which have different spatial resolution and detection surface. The unitcell of both arrays is a 3D cylindrical diode with a 25 [Formula: see text]m diameter and a 20 [Formula: see text]m depth that results in a welldefined and isolated radiation sensitive micro-volume etched inside a silicon wafer. Measurements were carried out at the Accélérateur Linéaire et Tandem à Orsay (ALTO) facility by irradiating the two detection systems with monoenergetic proton beams from 6 to 20 MeV at clinical-equivalent fluence rates. The microdosimetry quantities were obtained with a spatial resolution of 200 [Formula: see text]m and 600 [Formula: see text]m for the 11[Formula: see text]11 system and for the 3[Formula: see text]3 multi-array system, respectively. Experimental results were compared with Monte Carlo simulations and an overall good agreement was found. The good performance of both microdetector arrays demonstrates that this architecture and both configurations can be used clinically as microdosimeters for measuring the lineal energy distributions and, thus, for RBE optimization of hadron therapy treatments. Likewise, the results have shown that the devices can be also employed as a multipurpose device for beam monitoring in particle accelerators.

Verification of a Monte Carlo dose calculation engine in proton minibeam radiotherapy in a passive scattering beamline for preclinical trials.

OBJECTIVES: Proton minibeam radiation therapy (pMBRT) is a novel therapeutic strategy that combines the benefits of proton therapy with the remarkable normal tissue preservation observed with the use of submillimetric spatially fractionated beams. This promising technique has been implemented at the Institut Curie-Proton therapy centre (ICPO) using a first prototype of a multislit collimator. The purpose of this work was to develop a Monte Carlo-based dose calculation engine to reliably guide preclinical studies at ICPO. METHODS: The whole "Y1"-passive beamline at the ICPO, including pMBRT implementation, was modelled using the Monte Carlo GATE v. 7.0 code. A clinically relevant proton energy (100 MeV) was used as starting point. Minibeam generation by means of the brass collimator used in the first experiments was modelled. A virtual source was modelled at the exit of the beamline nozzle and outcomes were compared with dosimetric measurements performed with EBT3 gafchromic films and a diamond detector in water. Dose distributions were recorded in a water phantom and in rat CT images (7-week-old male Fischer rats). RESULTS: The dose calculation engine was benchmarked against experimental data and was then used to assess dose distributions in CT images of a rat, resulting from different irradiation configurations used in several experiments. It reduced computational time by an order of magnitude. This allows us to speed up simulations for in vivo trials, where we obtained peak-to-valley dose ratios of 1.20 ± 0.05 and 6.1 ± 0.2 for proton minibeam irradiations targeting the tumour and crossing the rat head. Tumour eradication was observed in the 67 and 22% of the animals treated respectively. CONCLUSION: A Monte Carlo dose calculation engine for pMBRT implementation with mechanical collimation has been developed. This tool can be used to guide and interpret the results of in vivo trials. ADVANCES IN KNOWLEDGE: This is the first Monte Carlo dose engine for pMBRT that is being used to guide preclinical trials in a clinical proton therapy centre.

Silicon 3D Microdetectors for Microdosimetry in Hadron Therapy.

The present overview describes the evolution of new microdosimeters developed in the National Microelectronics Center in Spain (IMB-CNM, CSIC), ranging from the first ultra-thin 3D diodes (U3DTHINs) to the advanced 3D cylindrical microdetectors, which have been developed over the last 10 years. In this work, we summarize the design, main manufacture processes, and electrical characterization of these devices. These sensors were specifically customized for use in particle therapy and overcame some of the technological challenges in this domain, namely the low noise capability, well-defined sensitive volume, high spatial resolution, and pile-up robustness. Likewise, both architectures reduce the loss of charge carriers due to trapping effects, the charge collection time, and the voltage required for full depletion compared to planar silicon detectors. In particular, a 3D‒cylindrical architecture with electrodes inserted into the silicon bulk and with a very well‒delimited sensitive volume (SV) mimicked a cell array with shapes and sizes similar to those of mammalian cells for the first time. Experimental tests of the carbon beamlines at the Grand Accélérateur National d'Lourds (GANIL, France) and Centro Nazionale Adroterapia Oncologica (CNAO, Italy) showed the feasibility of the U3DTHINs in hadron therapy beams and the good performance of the 3D‒cylindrical microdetectors for assessing linear energy distributions of clinical beams, with clinical fluence rates of 5 × 107 s-1cm-2 without saturation. The dose-averaged lineal energies showed a generally good agreement with Monte Carlo simulations. The results indicated that these devices can be used to characterize the microdosimetric properties in hadron therapy, even though the charge collection efficiency (CCE) and electronic noise may pose limitations on their performance, which is studied and discussed herein. In the last 3D‒cylindrical microdetector generation, we considerably improved the CCE due to the microfabrication enhancements, which have led to shallower and steeper dopant profiles. We also summarize the successive microdosimetric characterizations performed with both devices in proton and carbon beamlines.

Tumor Control in RG2 Glioma-Bearing Rats: A Comparison Between Proton Minibeam Therapy and Standard Proton Therapy.

PURPOSE: Proton minibeam radiation therapy (pMBRT) is a novel radiation therapy approach that exploits the synergies of proton therapy with the gain in normal tissue preservation observed upon irradiation with narrow, spatially fractionated, beams. The net gain in normal tissue sparing that has been shown by pMBRT may lead to the efficient treatment of very radioresistant tumors, which are currently mostly treated palliatively. The aim of this study was to perform an evaluation of the tumor effectiveness of proton minibeam radiation therapy for the treatment of RG2 glioma-bearing rats. METHODS AND MATERIALS: Two groups (n = 9) of RG2 glioma-bearing rats were irradiated with either standard proton therapy or with pMBRT, with a dose prescription of 25 Gy in 1 fraction. The animals were followed up for a maximum of 6 months. At the end of the study, histopathological studies were performed to assess both the tumor presence and the possible side effects. RESULTS: Tumor control was achieved in the 2 irradiated series, with superior survival in the pMBRT group compared with the standard proton therapy group. Long-term (>170 days) survival rates of 22% and 67% were obtained in the standard proton therapy and pMBRT groups, respectively. No tumor was observed in the histopathological analysis. Although animals with long-term survival in the standard radiation therapy exhibit substantial brain damage, including marked radionecrosis, less severe toxicity was observed in the pMBRT group. CONCLUSIONS: pMBRT offers a significant increase in the therapeutic index of brain tumors: The majority of the glioma-bearing rats (67%) survived 6 months with less severe side effects.

Effect of X-ray minibeam radiation therapy on clonogenic survival of glioma cells.

The goal is to compare, in vitro, the efficiency of minibeam radiotherapy (MBRT) and standard RT in inducing clonogenic cell death in glioma cell lines. With this aim, we report on the first in vitro study performed in an X-ray Small Animal Radiation Research Platform (SARRP) modified for minibeam irradiations. F98 rat and U87 human glioma cells were irradiated with either an array of minibeams (MB) or with conventional homogeneous beams (broad beam, BB). A specially designed multislit collimator was used to generate the minibeams with a with of a center-to-center distance of 1465 (±10) µm, and a PVDR value of 12.4 (±2.3) measured at 1 cm depth in a water phantom. Cells were either replated for clonogenic assay directly (immediate plating, IP) or 24 h after irradiation (delayed plating, DP) to assess the effect of potentially lethal damage repair (PLDR) on cell survival. Our hypothesis is that with MBRT, a similar level of clonogenic cell death can be reached compared to standard RT, when using equal mean radiation doses. To prove this, we performed dose escalations to determine the minimum integrated dose needed to reach a similar level of clonogenic cell death for both treatments. We show that this minimum dose can vary per cell line: in F98 cells a dose of 19 Gy was needed to obtain similar levels of clonogenic survival, whereas in U87 cells there was still a slightly increased survival with MB compared to BB 19 Gy treatment. The results suggest also an impairment of DNA damage repair in F98 cells as there is no difference in clonogenic cell survival between immediately and delayed plated cells for each dose and irradiation mode. For U87 cells, a small IP-DP effect was observed in the case of BB irradiation up to a dose of 17 Gy. However, at 19 Gy BB, as well as for the complete dose range of MB irradiation, U87 cells did not show a difference in clonogenic survival between IP and DP. We therefore speculate that MBRT might influence PLDR. The current results show that X-ray MBRT is a promising method for treatment of gliomas: future preclinical and clinical studies should aim at reaching a minimum radiation (valley) dose for effective eradication of gliomas with increased sparing of normal tissues compared to standard RT.

Implementation of planar proton minibeam radiation therapy using a pencil beam scanning system: A proof of concept study.

PURPOSE: Proton minibeam radiation therapy (pMBRT) is an innovative approach that combines the advantages of minibeam radiation therapy with the more precise ballistics of protons to further reduce the side effects of radiation. One of the main challenges of this approach is the generation of very narrow proton pencil beams with an adequate dose-rate to treat patients within a reasonable treatment time (several minutes) in existing clinical facilities. The aim of this study was to demonstrate the feasibility of implementing pMBRT by combining the pencil beam scanning (PBS) technique with the use of multislit collimators. This proof of concept study of pMBRT with a clinical system is intended to guide upcoming biological experiments. METHODS: Monte Carlo simulations (TOPAS v3.1.p2) were used to design a suitable multislit collimator to implement planar pMBRT for conventional pencil beam scanning settings. Dose distributions (depth-dose curves, lateral profiles, Peak-to-Valley Dose Ratio (PVDR) and dose-rates) for different proton beam energies were assessed by means of Monte Carlo simulations and experimental measurements in a water tank using commercial ionization chambers and a new p-type silicon diode, the IBA RAZOR. An analytical intensity-modulated dose calculation algorithm designed to optimize the weight of individual Bragg peaks composing the field was also developed and validated. RESULTS: Proton minibeams were then obtained using a brass multislit collimator with five slits measuring 2 cm × 400 µm in width with a center-to-center distance of 4 mm. The measured and calculated dose distributions (depth-dose curves and lateral profiles) showed a good agreement. Spread-out Bragg peaks (SOBP) and homogeneous dose distributions around the target were obtained by means of intensity modulation of Bragg peaks, while maintaining spatial fractionation at shallow depths. Mean dose-rates of 0.12 and 0.09 Gy/s were obtained for one iso-energy layer and a SOBP conditions in the presence of multislit collimator. CONCLUSIONS: This study demonstrates the feasibility of implementing pMBRT on a PBS system. It also confirms the reliability of RAZOR detector for pMBRT dosimetry. This newly developed experimental methodology will support the design of future preclinical research with pMBRT.

Proton minibeam radiation therapy widens the therapeutic index for high-grade gliomas.

Proton minibeam radiation therapy (pMBRT) is a novel strategy which has already shown a remarkable reduction in neurotoxicity as to compared with standard proton therapy. Here we report on the first evaluation of tumor control effectiveness in glioma bearing rats with highly spatially modulated proton beams. Whole brains (excluding the olfactory bulb) of Fischer 344 rats were irradiated. Four groups of animals were considered: a control group (RG2 tumor bearing rats), a second group of RG2 tumor-bearing rats and a third group of normal rats that received pMBRT (70 Gy peak dose in one fraction) with very heterogeneous dose distributions, and a control group of normal rats. The tumor-bearing and normal animals were followed-up for 6 months and one year, respectively. pMBRT leads to a significant tumor control and tumor eradication in 22% of the cases. No substantial brain damage which confirms the widening of the therapeutic window for high-grade gliomas offered by pMBRT. Additionally, the fact that large areas of the brain can be irradiated with pMBRT without significant side effects, would allow facing the infiltrative nature of gliomas.

Optimization of the mechanical collimation for minibeam generation in proton minibeam radiation therapy.

PURPOSE: The dose tolerances of normal tissues continue to be the main barrier in radiation therapy. To lower it, a novel concept based on a combination of proton therapy and the use of arrays of parallel and thin beams has been recently proposed: proton minibeam radiation therapy (pMBRT). It allies the inherent advantages of protons with the remarkable normal tissue preservation observed when irradiated with submillimetric spatially fractionated beams. Due to multiple Coulomb scattering, the tumor receives a homogeneous dose distribution, while normal tissues in the beam path benefit from the spatial fractionation of the dose. This promising technique has already been implemented at a clinical center (Proton therapy Center of Orsay) by means of a first prototype of a multislit collimator. The main goal of this work was to optimize the minibeam generation by means of a mechanical collimation. METHODS: Monte Carlo simulations (GATE V7.1) were used to evaluate the influence of the collimator material (brass, nickel, iron, tungsten), thickness, phantom-to-collimator distance (PCD), among other parameters, on the dose distributions. Maximization of the peak-to-valley dose ratios (PVDR) in normal tissues along with minimization of full width at half maximum, penumbras and neutron contamination were used as figures of merit. As a starting point for the optimization, the collimator employed in our previous works was used. It consisted in 400 µm × 2 cm slits with a center-to-center distance (c-t-c) of 3200 µm. As the main targets of pMBRT will be neurological cases, 100 MeV energy proton minibeams were considered. This energy range would allow treating tumors located at the center of the brain (the worst scenario). RESULTS: Tungsten and brass are the most advantageous materials among those considered. A tungsten collimator provides the highest PVDR and lowest penumbra. Although the neutron yield generated in the tungsten collimator is 3 times higher than that of the other materials, the biologic neutron doses at the patient position amount to less than 0.05% and 0.7% of the peak and valley doses, respectively. In addition, shorter PCD than the one currently used (7 cm) leads to thinner beams (enhancing the dose-volume effects), accompanied, however, by an increase of neutron dose at the phantom surface. Finally, no gain in dose distributions is obtained by using nonparallel slits. CONCLUSIONS: The collimator design and irradiation configuration have been optimized to minimize the angular spread, deliver the highest PVDR and the lowest valley possible in the normal tissues in pMBRT. We have also confirmed that even though the neutron yield generated in the multislit collimator is higher with respect to the one produced by the collimators used in conventional proton therapy, the increase of biological neutron dose in the patient will remain low (less than 1%).

Proton minibeam radiation therapy spares normal rat brain: Long-Term Clinical, Radiological and Histopathological Analysis.

Proton minibeam radiation therapy (pMBRT) is a novel strategy for minimizing normal tissue damage resulting from radiotherapy treatments. This strategy partners the inherent advantages of protons for radiotherapy with the gain in normal tissue preservation observed upon irradiation with narrow, spatially fractionated beams. In this study, whole brains (excluding the olfactory bulb) of Fischer 344 rats (n = 16) were irradiated at the Orsay Proton Therapy Center. Half of the animals received standard proton irradiation, while the other half were irradiated with pMBRT at the same average dose (25 Gy in one fraction). The animals were followed-up for 6 months. A magnetic resonance imaging (MRI) study using a 7-T small-animal MRI scanner was performed along with a histological analysis. Rats treated with conventional proton irradiation exhibited severe moist desquamation, permanent epilation and substantial brain damage. In contrast, rats in the pMBRT group exhibited no skin damage, reversible epilation and significantly reduced brain damage; some brain damage was observed in only one out of the eight irradiated rats. These results demonstrate that pMBRT leads to an increase in normal tissue resistance. This net gain in normal tissue sparing can lead to the efficient treatment of very radio-resistant tumours, which are currently mostly treated palliatively.