Evaluation of Radiation Sensitivity Differences in Mouse Liver Tumor Organoids Using CRISPR/Cas9-Mediated Gene Mutation.

BACKGROUND: To assess the radiosensitivity of liver tumors harboring different genetic mutations, mouse liver tumors were generated in vivo through the hydrodynamic injection of clustered regularly interspaced short palindromic repeat/caspase 9 (CRISPR/Cas9) constructs encoding single-guide RNAs (sgRNAs) targeting Tp53, Pten, Nf1, Nf2, Tsc2, Cdkn2a, or Rb1. METHODS: The plasmid vectors were delivered to the liver of adult C57BL/6 mice via hydrodynamic tail vein injection. The vectors were injected into 10 mice in each group. Organoids were generated from mouse liver tumors. The radiation response of the organoids was assessed using an ATP cell viability assay. RESULTS: The mean survival period of mice injected with vectors targeting Nf2 (4.8 months) was lower than that of other mice. Hematoxylin and eosin staining, immunohistochemical (IHC) staining, and target sequencing analyses revealed that mouse liver tumors harbored the expected mutations. Tumor organoids were established from mouse liver tumors. Histological evaluation revealed marked morphological similarities between the mouse liver tumors and the generated tumor organoids. Moreover, IHC staining indicated that the parental tumor protein expression pattern was maintained in the organoids. The results of the ATP cell viability assay revealed that the tumor organoids with mutated Nf2 were more resistant to high-dose radiation than those with other gene mutations. CONCLUSIONS: This study developed a radiation response assessment system for mouse tumors with mutant target genes using CRISPR/Cas9 and organoids. The Tp53 and Pten double mutation in combination with the Nf2 mutation increased the radiation resistance of tumors. The system used in this study can aid in elucidating the mechanism underlying differential intrinsic radiation sensitivity of individual tumors.

Optical Filter-Embedded Fiber-Optic Radiation Sensor for Ultra-High Dose Rate Electron Beam Dosimetry.

FLASH radiotherapy is an emerging radiotherapy technique used to spare normal tissues. It employs ultra-high dose rate radiation beams over 40 Gy/s, which is significantly higher than those of conventional radiotherapy. In this study, a fiber-optic radiation sensor (FORS) was fabricated using a plastic scintillator, an optical filter, and a plastic optical fiber to measure the ultra-high dose rate electron beams over 40 Gy/s used in FLASH radiotherapy. The radiation-induced emissions, such as Cherenkov radiation and fluorescence generated in a transmitting optical fiber, were spectrally discriminated from the light outputs of the FORS. To evaluate the linearity and dose rate dependence of the FORS, the outputs of the fiber-optic radiation sensor were measured according to distances from an electron scattering device, and the results were compared with those of an ionization chamber and radiochromic films. Finally, the percentage depth doses were obtained using the FORS as a function of depth in a water phantom. This study found that ultra-high dose rate electron beams over 40 Gy/s could be measured in real time using a FORS.

Feasibility of Ultra-Thin Fiber-Optic Dosimeters for Radiotherapy Dosimetry.

In this study, prototype ultra-thin fiber-optic dosimeters were fabricated using organic scintillators, wavelength shifting fibers, and plastic optical fibers. The sensor probes of the ultra-thin fiber-optic dosimeters consisted of very thin organic scintillators with thicknesses of 100, 150 and 200 µm. These types of sensors cannot only be used to measure skin or surface doses but also provide depth dose measurements with high spatial resolution. With the ultra-thin fiber-optic dosimeters, surface doses for gamma rays generated from a Co-60 therapy machine were measured. Additionally, percentage depth doses in the build-up regions were obtained by using the ultra-thin fiber-optic dosimeters, and the results were compared with those of external beam therapy films and a conventional fiber-optic dosimeter.

Performance Evaluation of a Multichannel All-In-One Phantom Dosimeter for Dose Measurement of Diagnostic X-ray Beam.

We developed a multichannel all-in-one phantom dosimeter system composed of nine sensing probes, a chest phantom, an image intensifier, and a complementary metal-oxide semiconductor (CMOS) image sensor to measure the dose distribution of an X-ray beam used in radiation diagnosis. Nine sensing probes of the phantom dosimeter were fabricated identically by connecting a plastic scintillating fiber (PSF) to a plastic optical fiber (POF). To measure the planar dose distribution on a chest phantom according to exposure parameters used in clinical practice, we divided the top of the chest phantom into nine equal parts virtually and then installed the nine sensing probes at each center of the nine equal parts on the top of the chest phantom as measuring points. Each scintillation signal generated in the nine sensing probes was transmitted through the POFs and then intensified by the image intensifier because the scintillation signal normally has a very low light intensity. Real-time scintillation images (RSIs) containing the intensified scintillation signals were taken by the CMOS image sensor with a single lens optical system and displayed through a software program. Under variation of the exposure parameters, we measured RSIs containing dose information using the multichannel all-in-one phantom dosimeter and compared the results with the absorbed doses obtained by using a semiconductor dosimeter (SCD). From the experimental results of this study, the light intensities of nine regions of interest (ROI) in the RSI measured by the phantom dosimeter were similar to the dose distribution obtained using the SCD. In conclusion, we demonstrated that the planar dose distribution including the entrance surface dose (ESD) can be easily measured by using the proposed phantom dosimeter system.

Development of a Small-Sized, Flexible, and Insertable Fiber-Optic Radiation Sensor for Gamma-Ray Spectroscopy.

We fabricated a small-sized, flexible, and insertable fiber-optic radiation sensor (FORS) that is composed of a sensing probe, a plastic optical fiber (POF), a photomultiplier tube (PMT)-amplifier system, and a multichannel analyzer (MCA) to obtain the energy spectra of radioactive isotopes. As an inorganic scintillator for gamma-ray spectroscopy, a cerium-doped lutetium yttrium orthosilicate (LYSO:Ce) crystal was used and two solid-disc type radioactive isotopes with the same dimensions, cesium-137 (Cs-137) and cobalt-60 (Co-60), were used as gamma-ray emitters. We first determined the length of the LYSO:Ce crystal considering the absorption of charged particle energy and measured the gamma-ray energy spectra using the FORS. The experimental results demonstrated that the proposed FORS can be used to discriminate species of radioactive isotopes by measuring their inherent energy spectra, even when gamma-ray emitters are mixed. The relationship between the measured photon counts of the FORS and the radioactivity of Cs-137 was subsequently obtained. The amount of scintillating light generated from the FORS increased by increasing the radioactivity of Cs-137. Finally, the performance of the fabricated FORS according to the length and diameter of the POF was also evaluated. Based on the results of this study, it is anticipated that a novel FORS can be developed to accurately measure the gamma-ray energy spectrum in inaccessible locations such as narrow areas and holes.

Development of a novel proton dosimetry system using an array of fiber-optic Cerenkov radiation sensors.

This study describes the development and evaluation of a new dosimetric system for proton therapy using an array of fiber-optic Cerenkov radiation sensors (AFCRS). The AFCRS was superior to a conventional, multi-layer ion chamber (MLIC) system in real-time data acquisition and cost effectiveness.

Effects of Temperature and X-rays on Plastic Scintillating Fiber and Infrared Optical Fiber.

In this study, we have studied the effects of temperature and X-ray energy variations on the light output signals from two different fiber-optic sensors, a fiber-optic dosimeter (FOD) based on a BCF-12 as a plastic scintillating fiber (PSF) and a fiber-optic thermometer (FOT) using a silver halide optical fiber as an infrared optical fiber (IR fiber). During X-ray beam irradiation, the scintillating light and IR signals were measured simultaneously using a dosimeter probe of the FOD and a thermometer probe of the FOT. The probes were placed in a beaker with water on the center of a hotplate, under variation of the tube potential of a digital radiography system or the temperature of the water in the beaker. From the experimental results, in the case of the PSF, the scintillator light output at the given tube potential decreased as the temperature increased in the temperature range from 25 to 60 °C. We demonstrated that commonly used BCF-12 has a significant temperature dependence of -0.263 ± 0.028%/°C in the clinical temperature range. Next, in the case of the IR fiber, the intensity of the IR signal was almost uniform at each temperature regardless of the tube potential range from 50 to 150 kVp. Therefore, we also demonstrated that the X-ray beam with an energy range used in diagnostic radiology does not affect the IR signals transmitted via a silver halide optical fiber.

A fiber-optic sensor using an aqueous solution of sodium chloride to measure temperature and water level simultaneously.

A fiber-optic sensor system using a multiplexed array of sensing probes based on an aqueous solution of sodium chloride (NaCl solution) and an optical time-domain reflectometer (OTDR) for simultaneous measurement of temperature and water level is proposed. By changing the temperature, the refractive index of the NaCl solution is varied and Fresnel reflection arising at the interface between the distal end of optical fiber and the NaCl solution is then also changed. We measured the modified optical power of the light reflected from the sensing probe using a portable OTDR device and also obtained the relationship between the temperature of water and the optical power. In this study, the water level was simply determined by measuring the signal difference of the optical power due to the temperature difference of individual sensing probes placed inside and outside of the water. In conclusion, we demonstrate that the temperature and water level can be obtained simultaneously by measuring optical powers of light reflected from sensing probes based on the NaCl solution. It is anticipated that the proposed fiber-optic sensor system makes it possible to remotely monitor the real-time change of temperature and water level of the spent fuel pool during a loss of power accident.

The addition of mifamurtide to chemotherapy improves lifetime effectiveness in children with osteosarcoma: a Markov model analysis.

In the absence of long-term clinical trials that compare mifamurtide plus chemotherapy versus chemotherapy only for treatment of osteosarcoma, decision analysis is a useful tool that helps to determine the optimal treatment strategy. We analyzed the differences between mifamurtide plus chemotherapy versus chemotherapy only by using modeling to determine the treatment approach that results in longer life expectancy among children with osteosarcoma. We used the Markov model to compare the expected lifetime quality-adjusted life years (QALYs) between mifamurtide plus chemotherapy versus chemotherapy only. Our target cohort consisted of children with osteosarcoma. The starting age of the cohort was 12 years and cycle length was 3 months. The transition probabilities for each disease state and death were calculated using overall survival or progression free survival data from randomized controlled trials. Utility weights from scenario-based survey for 303 Korean general populations were applied to the model. Based on the base case analysis, the incremental benefit analysis indicated that mifamurtide plus chemotherapy resulted in an incremental QALY increase of 1.57 (a relative increase of 16.3 % in QALY expectancy) compared to chemotherapy only. Also, the incremental life years gained (LYG) from mifamurtide plus chemotherapy was 1.96 on comparison with chemotherapy only; this is a relative increase of 15.7 % in LYG expectancy. The decision analysis model indicated that mifamurtide plus chemotherapy was associated with a substantially longer survival than chemotherapy only among children with osteosarcoma during their lifetime.

Measurement of Cerenkov radiation induced by the gamma-rays of Co-60 therapy units using wavelength shifting fiber.

In this study, a wavelength shifting fiber that shifts ultra-violet and blue light to green light was employed as a sensor probe of a fiber-optic Cerenkov radiation sensor. In order to characterize Cerenkov radiation generated in the developed wavelength shifting fiber and a plastic optical fiber, spectra and intensities of Cerenkov radiation were measured with a spectrometer. The spectral peaks of light outputs from the wavelength shifting fiber and the plastic optical fiber were measured at wavelengths of 500 and 510 nm, respectively, and the intensity of transmitted light output of the wavelength shifting fiber was 22.2 times higher than that of the plastic optical fiber. Also, electron fluxes and total energy depositions of gamma-ray beams generated from a Co-60 therapy unit were calculated according to water depths using the Monte Carlo N-particle transport code. The relationship between the fluxes of electrons over the Cerenkov threshold energy and the energy depositions of gamma-ray beams from the Co-60 unit is a near-identity function. Finally, percentage depth doses for the gamma-ray beams were obtained using the fiber-optic Cerenkov radiation sensor, and the results were compared with those obtained by an ionization chamber. The average dose difference between the results of the fiber-optic Cerenkov radiation sensor and those of the ionization chamber was about 2.09%.

Measurement of entrance surface dose on an anthropomorphic thorax phantom using a miniature fiber-optic dosimeter.

A miniature fiber-optic dosimeter (FOD) system was fabricated using a plastic scintillating fiber, a plastic optical fiber, and a multi-pixel photon counter to measure real-time entrance surface dose (ESD) during radiation diagnosis. Under varying exposure parameters of a digital radiography (DR) system, we measured the scintillating light related to the ESD using the sensing probe of the FOD, which was placed at the center of the beam field on an anthropomorphic thorax phantom. Also, we obtained DR images using a flat panel detector of the DR system to evaluate the effects of the dosimeter on image artifacts during posteroanterior (PA) chest radiography. From the experimental results, the scintillation output signals of the FOD were similar to the ESDs including backscatter simultaneously obtained using a semiconductor dosimeter. We demonstrated that the proposed miniature FOD can be used to measure real-time ESDs with minimization of DR image artifacts in the X-ray energy range of diagnostic radiology.

Feasibility of fiber-optic radiation sensor using Cerenkov effect for detecting thermal neutrons.

In this research, we propose a novel method for detecting thermal neutrons with a fiber-optic radiation sensor using the Cerenkov effect. We fabricate a fiber-optic radiation sensor that detects thermal neutrons with a Gd-foil, a rutile crystal, and a plastic optical fiber. The relationship between the fluxes of electrons inducing Cerenkov radiation in the sensor probe of the fiber-optic radiation sensor and thermal neutron fluxes is determined using the Monte Carlo N-particle transport code simulations. To evaluate the fiber-optic radiation sensor, the Cerenkov radiation generated in the fiber-optic radiation sensor by irradiation of pure thermal neutron beams is measured according to the depths of polyethylene.

Application of Cerenkov radiation generated in plastic optical fibers for therapeutic photon beam dosimetry.

A Cerenkov fiber-optic dosimeter (CFOD) is fabricated using plastic optical fibers to measure Cerenkov radiation induced by a therapeutic photon beam. We measured the Cerenkov radiation generated in optical fibers in various irradiation conditions to evaluate the usability of Cerenkov radiation for a photon beam therapy dosimetry. As a results, the spectral peak of Cerenkov radiation was measured at a wavelength of 515 nm, and the intensity of Cerenkov radiation increased linearly with increasing irradiated length of the optical fiber. Also, the intensity peak of Cerenkov radiation was measured in the irradiation angle range of 30 to 40 deg. In the results of Monte Carlo N-particle transport code simulations, the relationship between fluxes of electrons over Cerenkov threshold energy and energy deposition of a 6 MV photon beam had a nearly linear trend. Finally, percentage depth doses for the 6 MV photon beam could be obtained using the CFOD and the results were compared with those of an ionization chamber. Here, the mean dose difference was about 0.6%. It is anticipated that the novel and simple CFOD can be effectively used for measuring depth doses in radiotherapy dosimetry.

Simultaneous measurements of pure scintillation and Cerenkov signals in an integrated fiber-optic dosimeter for electron beam therapy dosimetry.

For real-time dosimetry in electron beam therapy, an integrated fiber-optic dosimeter (FOD) is developed using a water-equivalent dosimeter probe, four transmitting optical fibers, and a multichannel light-measuring device. The dosimeter probe is composed of two inner sensors, a scintillation sensor and a Cerenkov sensor, and each sensor has two different channels. Accordingly, we measured four separate light signals from each channel in the dosimeter probe, simultaneously, and then obtained the scintillation and Cerenkov signals using a subtraction method. To evaluate the performance of the integrated FOD, we measured the light signals according to the irradiation angle of the electron beam, the depth variation of the solid water phantom, and the electron beam energy. In conclusion, we demonstrated that the pure scintillation and Cerenkov signals obtained by an integrated FOD system based on a subtraction method can be effectively used for calibrating the conditions of high-energy electron beams in radiotherapy.

Water-equivalent one-dimensional scintillating fiber-optic dosimeter for measuring therapeutic photon beam.

In this study, we fabricated a one-dimensional scintillating fiber-optic dosimeter, which consists of 9 scintillating fiber-optic dosimeters, septa, and PMMA blocks for measuring surface and percentage depth doses of a therapeutic photon beam. Each dosimeter embedded in the 1-D scintillating fiber-optic dosimeter is composed of square type organic scintillators and plastic optical fibers. Also black PVC films are used as septa to minimize cross-talk between the scintillating fiber-optic dosimeters. To construct a dosimeter system, a 1-D scintillating fiber-optic dosimeter and a CMOS image sensor were combined with 20 m-length plastic optical fibers. Using the dosimeter system, we measured surface and percentage depth doses of 6 and 15 MV photon beams and compared the results with those of EBT films and an ionization chamber.

Integral T-shaped phantom-dosimeter system to measure transverse and longitudinal dose distributions simultaneously for stereotactic radiosurgery dosimetry.

A T-shaped fiber-optic phantom-dosimeter system was developed using square scintillating optical fibers, a lens system, and a CMOS image camera. Images of scintillating light were used to simultaneously measure the transverse and longitudinal distributions of absorbed dose of a 6 MV photon beam with field sizes of 1 × 1 and 3 × 3 cm(2). Each optical fiber has a very small sensitive volume and the sensitive material is water equivalent. This allows the measurements of cross-beam profile as well as the percentage depth dose of small field sizes. In the case of transverse dose distribution, the measured beam profiles were gradually become uneven and the beam edge had a gentle slope with increasing depth of the PMMA phantom. In addition, the maximum dose values of longitudinal dose distribution for 6 MV photon beam with field sizes of 1 × 1 and 3 × 3 cm(2) were found to be at a depth of approximately 15 mm and the percentage depth dose of both field sizes were nearly in agreement at the skin dose level. Based on the results of this study, it is anticipated that an all-in-one phantom-dosimeter can be developed to accurately measure beam profiles and dose distribution in a small irradiation fields prior to carrying out stereotactic radiosurgery.

Fiber-optic Cerenkov radiation sensor for proton therapy dosimetry.

In proton therapy dosimetry, a fiber-optic radiation sensor incorporating a scintillator must undergo complicated correction processes due to the quenching effect of the scintillator. To overcome the drawbacks of the fiber-optic radiation sensor, we proposed an innovative method using the Cerenkov radiation generated in plastic optical fibers. In this study, we fabricated a fiber-optic Cerenkov radiation sensor without an organic scintillator to measure Cerenkov radiation induced by therapeutic proton beams. Bragg peaks and spread-out Bragg peaks of proton beams were measured using the fiber-optic Cerenkov radiation sensor and the results were compared with those of an ionization chamber and a fiber-optic radiation sensor incorporating an organic scintillator. From the results, we could obtain the Bragg peak and the spread-out Bragg peak of proton beams without quenching effects induced by the scintillator, and these results were in good agreement with those of the ionization chamber. We also measured the Cerenkov radiation generated from the fiber-optic Cerenkov radiation sensor as a function of the dose rate of the proton beam.

Measurements of relative depth doses and Cerenkov light using a scintillating fiber-optic dosimeter with Co-60 radiotherapy source.

In this study, we fabricated a scintillating fiber-optic dosimeter, which consists of an organic scintillator and a plastic optical fiber, for radiotherapy dosimetry. To select an adequate kind and length of scintillator for γ-rays generated from a Co-60 source, scintillating light from various kinds and lengths of organic scintillators is measured. Using a scintillating fiber-optic dosimeter, the γ-rays generated from a Co-60 therapy unit are measured and relative doses are obtained according to the field size of the γ-ray beam and the depth in a water phantom. Also, Cerenkov light generated by the interactions of primary or secondary electrons and the plastic optical fiber is measured with different field sizes and depths of a water phantom using a background optical fiber.

Development of a 2-channel embedded infrared fiber-optic temperature sensor using silver halide optical fibers.

A 2-channel embedded infrared fiber-optic temperature sensor was fabricated using two identical silver halide optical fibers for accurate thermometry without complicated calibration processes. In this study, we measured the output voltages of signal and reference probes according to temperature variation over a temperature range from 25 to 225 °C. To decide the temperature of the water, the difference between the amounts of infrared radiation emitted from the two temperature sensing probes was measured. The response time and the reproducibility of the fiber-optic temperature sensor were also obtained. Thermometry with the proposed sensor is immune to changes if parameters such as offset voltage, ambient temperature, and emissivity of any warm object. In particular, the temperature sensing probe with silver halide optical fibers can withstand a high temperature/pressure and water-chemistry environment. It is expected that the proposed sensor can be further developed to accurately monitor temperature in harsh environments.

Development and characterization of the integrated fiber-optic radiation sensor for the simultaneous detection of neutrons and gamma rays.

Sometimes, detection of thermal neutrons in the presence of gamma rays is required. This study developed and characterized an integrated fiber-optic radiation sensor for the simultaneous detection of thermal neutrons and gamma rays in a mixed radiation field. The performance of the integrated sensor was verified by measuring the distributions of thermal neutrons and gamma rays released from a nuclear fuel rod at the Kyoto University Critical Assembly. The experimental results show that the integrated sensor produced similar distribution patterns to those of thermal neutrons and gamma rays released from a fuel rod.