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This Roadmap paper covers the field of precision preclinical x-ray radiation studies in animal models. It is mostly focused on models for cancer and normal tissue response to radiation, but also discusses other disease models. The recent technological evolution in imaging, irradiation, dosimetry and monitoring that have empowered these kinds of studies is discussed, and many developments in the near future are outlined. Finally, clinical translation and reverse translation are discussed., (Creative Commons Attribution license.)

Purpose: To determine the dosimetric impact of using unedited autocontours in daily plan adaptation of patients with locally advanced pancreatic cancer (LAPC) treated with stereotactic body radiotherapy using tumor tracking., Materials and Methods: The study included 98 daily CT scans of 35 LAPC patients. All scans were manually contoured (MAN), and included the PTV and main organs-at-risk (OAR): stomach, duodenum and bowel. Precision and MIM deformable image registration (DIR) methods followed by contour propagation were used to generate autocontour sets on the daily CT scans. Autocontours remained unedited, and were compared to MAN on the whole organs and at 3, 1 and 0.5 cm from the PTV. Manual and autocontoured OAR were used to generate daily plans using the VOLO™ optimizer, and were compared to non-adapted plans. Resulting planned doses were compared based on PTV coverage and OAR dose-constraints., Results: Overall, both algorithms reported a high agreement between unclipped MAN and autocontours, but showed worse results when being evaluated on the clipped structures at 1 cm and 0.5 cm from the PTV. Replanning with unedited autocontours resulted in better OAR sparing than non-adapted plans for 95% and 84% plans optimized using Precision and MIM autocontours, respectively, and obeyed OAR constraints in 64% and 56% of replans., Conclusion: For the majority of fractions, manual correction of autocontours could be avoided or be limited to the region closest to the PTV. This practice could further reduce the overall timings of adaptive radiotherapy workflows for patients with LAPC., Competing Interests: All authors are employed by the Erasmus MC. MM and MH report serving as an advisory board member for Accuray during the conduct of the study., (Copyright © 2022 Magallon-Baro, Milder, Granton, Toom, Nuyttens and Hoogeman.)

Importance: Palliative thoracic radiotherapy (RT) can alleviate local symptoms associated with advanced non-small cell lung cancer (NSCLC), but esophagitis is a common treatment-related adverse event. Whether esophageal-sparing intensity-modulated RT (ES-IMRT) achieves a clinically relevant reduction in esophageal symptoms remains unclear., Objective: To examine whether ES-IMRT achieves a clinically relevant reduction in esophageal symptoms compared with standard RT., Design, Setting, and Participants: Palliative Radiation for Advanced Central Lung Tumors With Intentional Avoidance of the Esophagus (PROACTIVE) is a multicenter phase 3 randomized clinical trial that enrolled patients between June 24, 2016, and March 6, 2019. Data analysis was conducted from January 23, 2020, to October 22, 2021. Patients had up to 1 year of follow-up. Ninety patients at 6 tertiary academic cancer centers who had stage III/IV NSCLC and were eligible for palliative thoracic RT (20 Gy in 5 fractions or 30 Gy in 10 fractions) were included., Interventions: Patients were randomized (1:1) to standard RT (control arm) or ES-IMRT. Target coverage was compromised to ensure the maximum esophagus dose was no more than 80% of the RT prescription dose., Main Outcomes and Measures: The primary outcome was esophageal quality of life (QOL) 2 weeks post-RT, measured by the esophageal cancer subscale (ECS) of the Functional Assessment of Cancer Therapy: Esophagus questionnaire. Higher esophageal cancer subscale scores correspond with improved QOL, with a 2- to 3-point change considered clinically meaningful. Secondary outcomes included overall survival, toxic events, and other QOL metrics. Intention-to-treat analysis was used., Results: Between June 24, 2016, and March 6, 2019, 90 patients were randomized to standard RT or ES-IMRT (median age at randomization, 72.0 years [IQR, 65.6-80.3]; 50 [56%] were female). Thirty-six patients (40%) received 20 Gy and 54 (60%) received 30 Gy. For the primary end point, the mean (SD) 2-week ECS score was 50.5 (10.2) in the control arm (95% CI, 47.2-53.8) and 54.3 (7.6) in the ES-IMRT arm (95% CI, 51.9-56.7) (P = .06). Symptomatic RT-associated esophagitis occurred in 24% (n = 11) of patients in the control arm vs 2% (n = 1) in the ES-IMRT arm (P = .002). In a post hoc subgroup analysis based on the stratification factor, reduction in esophagitis was most evident in patients receiving 30 Gy (30% [n = 8] vs 0%; P = .004). Overall survival was similar with standard RT (median, 8.6; 95% CI, 5.7-15.6 months) and ES-IMRT (median, 8.7; 95% CI, 5.1-10.2 months) (P = .62)., Conclusions and Relevance: In this phase 3 randomized clinical trial, ES-IMRT did not significantly improve esophageal QOL but significantly reduced the incidence of symptomatic esophagitis. Because post hoc analysis found that reduced esophagitis was most evident in patients receiving 30 Gy of RT, these findings suggest that ES-IMRT may be most beneficial when the prescription dose is higher (30 Gy)., Trial Registration: ClinicalTrials.gov Identifier: NCT02752126.

Purpose: To study the trade-offs of three online strategies to adapt treatment plans of patients with locally advanced pancreatic carcinoma (LAPC) treated using the CyberKnife with tumor tracking., Methods and Materials: A total of 35 planning computed tomography scans and 98 daily in-room computed tomography scans were collected from 35 patients with LAPC. Planned dose distributions, optimized with VOLO, were evaluated on manually contoured daily anatomies to collect daily doses. Three strategies were tested to adapt treatment plans: (1) unrestricted full replanning using a patient-specific plan template, (2) time-restricted replanning on organs at risk (OARs) within 3 cm from the planning target volume (PTV) structure, and (3) dose realignment optimization to stay within OAR constraints. Dose distributions resulting from each plan adaptation strategy were dosimetrically compared by means of gross tumor volume (GTV), PTV coverage, and OAR tolerances., Results: Planned doses did not result in dose-constraint violations for 28 of 98 daily anatomies. None of the suggested plan adaptation strategies improved planned doses significantly for this subset. For 70 of the 98 reported violations, the median (interquartile range) PTV coverage of the planned dose was 84% (76% to 86%). After plan adaptation, unrestricted replanning achieved clinically acceptable plans in 93% of these fractions, time-restricted replanning in 90%, and dose realignment in 74%, at median computational times of 8.5, 3, and 0.5 minutes. Over all 98 fractions, PTV coverage was reduced: -1% (-3% to 1%), -2% (-5% to 0%), and -2% (-8% to 0%) after each strategy, respectively. In 3 of 70 fractions, none of the suggested strategies achieved clinically acceptable OAR dose volumes., Conclusions: Unrestricted replanning was the most time-consuming method but reached the highest number of successfully adapted plans. Time-restricted replanning and dose realignment resulted in a high number of plans within dose constraints. Depending on the resources available, an adaptive strategy can be selected for each patient to address the specific anatomic challenges on the treatment day. The increase in the complexity of the strategy corresponds with an increasing number of successfully adapted plans., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background : Stereotactic body radiation therapy (SBRT) results in high local control (LC) rates in patients with non-small cell lung cancer (NSCLC). For central lung tumors, risk-adapted fractionation schedules are used and underdosage to the Planned Target Volume (PTV) is often accepted to respect the dose constraints of the organs at risk in order to avoid high rates of toxicity. The purpose of this study was to analyze the effect of PTV underdosage and other possible prognostic factors on local- and disease control after SBRT in patients with central lung tumors. Material and Methods : Patients with centrally located NSCLC treated with SBRT were included. The doses were converted into biologically equivalent dose using α/β-value of 10 Gy (BED 10 ). Underdosage to the PTV was defined as the (percentage of) PTV receiving less than 100 Gy BED 10 ; (%)PTV < 100 BED 10 . Potential prognostic factors for LC and Disease Free Survival (DFS) were evaluated using Cox regression analysis. Results : Two hundred and twenty patients received ≤12 fractions of SBRT. LC-rates were 88% at 2 years and 81% at 3 years. Twenty-seven patients developed a local recurrence. Both the PTV < 100 BED 10 and %PTV < 100 BED 10 were not prognostic for LC. Tumor size and forced expiratory volume in 1 second (FEV 1 ) were independently prognostic for LC. Disease progression was reported in 75 patients with DFS-rates of 66% at 2 years and 56% at 3 years. Disease recurrence was independent significantly associated with larger tumor diameter, lower lobe tumor location and decreased FEV 1 . Grade 4-5 toxicity was reported in 10 patients (8 with ultra-central tumors) and was fatal in at least 3 patients. Conclusion : Decrease in tumor coverage was not correlated with the local recurrence probability. The LC and DFS were promising after SBRT of centrally located NSCLC with tumor size, FEV 1 and tumor location (for DFS only) as prognostic factors.

Purpose: Locally advanced pancreatic cancer (LAPC) patients are prone to experience daily anatomical variations, which can lead to additional doses in organs-at-risk (OAR) during SBRT. A patient selection tool was developed to identify who may be at risk of exceeding dose tolerances, by quantifying the dosimetric impact of daily variations using an OAR motion model., Materials and Methods: The study included 133 CT scans from 35 LAPC patients. By following a leave-one-out approach, an OAR motion model trained with the remaining 34 subjects variations was used to simulate organ deformations on the left-out patient planning CT anatomy. Dose-volume histograms obtained from planned doses sampled on simulated organs resulted in the probability of exceeding OAR dose-constraints due to anatomical variations. Simulated probabilities were clustered with a threshold per organ according to clinical observations. If the prediction of at least one OAR was above the established thresholds, the patient was classified as being at risk., Results: Clinically, in 20/35 patients at least one OAR exceeded dose-constraints in the daily CTs. The model-based prediction had an accuracy of 89%, 71%, 91% in estimating the risk of exceeding dose tolerances for the duodenum, stomach and bowel, respectively. By combining the three predictions, our approach resulted in a correct patient classification for 29/35 patients (83%) when compared with clinical observations., Conclusions: Our model-based patient selection tool is able to predict who might be at risk of exceeding dose-constraints during SBRT. It is a promising tool to tailor LAPC treatments, e.g. by employing online adaptive SBRT; and hence, to minimize toxicity of patients being at risk., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Purpose: To characterize daily geometrical variations of gastrointestinal organs with respect to pancreatic tumors, through a population-based statistical model., Materials and Methods: The study included 131 CT scans from 35 pancreatic cancer patients treated with Stereotactic Body Radiotherapy (SBRT). For each patient, day-to-day anatomical variations of the stomach, the duodenum and the bowel were assessed from the deformation vector fields (DVF) obtained by non-rigidly registering the contours of the fractions to the planning CT scans. For the whole population, day-to-day motion-deformation patterns were abstracted using principal component analysis (PCA) on the set of DVFs mapped on a reference patient. Based on these geometrical variations, anatomies were generated to create population-based dose-volume histograms (DVH) per patient, which were also compared to clinical values., Results: Through PCA, the most dominant directions of daily deformations were localized in the abdominal organs. Common patterns were found, such as stomach contraction-expansion in the anterior-posterior direction ranging from 5 to 13 mm, and superior-inferior deformations on the bowel from 7 to 14 mm. The duodenum resulted to move laterally, but in a lesser extent (4-8 mm). The population-based DVHs derived from the model mostly included the daily DVHs observed in the clinic (in >90% of the cases)., Conclusions: Anatomical variations influence the delivered doses to healthy organs during SBRT. A motion model was successfully built and explored to extract the larger directions of movement of the gastrointestinal organs. Day-to-day motion modeling can potentially be used to account for geometrical uncertainties in future plan optimization and in online adaptive strategies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Stereotactic body radiotherapy (SBRT) has emerged as an effective option in oligo-metastatic cancer patients affected by lymph node metastases, but its use might be questioned due to risk of regional and distant dissemination through the lymph node chain. The primary aim of our study was to assess the loco-regional control following SBRT in this setting. Ninety-one patients undergoing SBRT for at least one lymph node metastasis from miscellaneous primary tumors were retrospectively evaluated for patterns of failure and toxicity. locoregional relapse-free survival (LRRFS) and distant metastasis-free survival (DMFS) at 4 years were 79 and 44%. Repeated use of local therapy after progression resulted in a median interval of 17 months until allocation to systemic therapy or supportive care. Forty-three percent of patients were alive at 4 years. Local failure, occurring in 15% of patients, was the only predictor of poor survival (HR: 3.06). Tumor diameter ≥ 30 mm and urothelial primary tumor predicted for impaired local control (HR: 4.59 and 5.43, respectively). Metastases from pulmonary cancer showed a significant earlier distant dissemination (HR: 3.53). Only acute and late grade 1-2 toxicities were reported except for 1 case of G3 dysphagia. Loco-regional failure risk is low (18%) and justifies the use of local therapies for patients with oligometastatic disease. Durable disease remission can be achieved by iterative use of local approaches. Local control is correlated to improved OS. Diameter and primary tumor type may affect response to SBRT and risk for early metastatic dissemination.

Background: Proton therapy is becoming increasingly available, so it is important to apply objective and individualized patient selection to identify those who are expected to benefit most from proton therapy compared to conventional intensity modulated radiation therapy (IMRT). Comparative treatment planning using normal tissue complication probability (NTCP) evaluation has recently been proposed. This work investigates the impact of NTCP model and dose uncertainties on model-based patient selection., Material and Methods: We used IMRT and intensity modulated proton therapy (IMPT) treatment plans of 78 oropharyngeal cancer patients, which were generated based on automated treatment planning and evaluated based on three published NTCP models. A reduction in NTCP of more than a certain threshold (e.g. 10% lower NTCP) leads to patient selection for IMPT, referred to as 'nominal' selection. To simulate the effect of uncertainties in NTCP-model coefficients (based on reported confidence intervals) and planned doses on the accuracy of model-based patient selection, the Monte Carlo method was used to sample NTCP-model coefficients and doses from a probability distribution centered at their nominal values. Patient selection accuracy within a certain sample was defined as the fraction of patients which had similar selection in both the 'nominal' and 'sampled' scenario., Results: For all three NTCP models, the median patient selection accuracy was found to be above 70% when only NTCP-model uncertainty was considered. Selection accuracy decreased with increasing uncertainty resulting from differences between planned and delivered dose. In case of excessive dose uncertainty, selection accuracy decreased to 60%., Conclusion: Model and dose uncertainty highly influence the accuracy of model-based patient selection for proton therapy. A reduction of NTCP-model uncertainty is necessary to reach more accurate model-based patient selection.

Background: Nintedanib has anti-fibrotic and anti-inflammatory activity and is approved for the treatment of idiopathic pulmonary fibrosis. The aim of this study was to noninvasively assess the efficacy of nintedanib in a mouse model of partial lung irradiation to prevent radiation-induced lung damage (RILD)., Methods: 266 C57BL/6 adult male mice were irradiated with a single radiation dose (0, 4, 8, 12, 16 or 20Gy) using parallel-opposed fields targeting the upper right lung using a precision image-guided small animal irradiator sparing heart and spine based on micro-CT images. One week post irradiation, mice were randomized across nintedanib daily oral gavage treatment (0, 30 or 60mg/kg). CT density analysis of the lungs was performed on monthly acquired micro-CT images. After 39weeks, lungs were processed to evaluate the fibrotic phenotype., Results: Although the CT density increase correlated with the radiation dose, nintedanib did not influence this relationship. Immunohistochemical analysis confirmed the ability of nintedanib to reduce the microscopic fibrotic phenotype, in particular interstitial edema, interstitial and perivascular fibrosis and inflammation, and vasculitis., Conclusions: Nintedanib reduces radiation-induced lung fibrosis after partial lung irradiation without adverse effects, however, noninvasive CT imaging measuring electron density cannot be applied for monitoring its effects., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Palliative thoracic radiotherapy is an effective technique to alleviate symptoms of disease burden in advanced-stage lung cancer patients. Previous randomized controlled studies demonstrated a survival benefit in patients with good performance status at radiation doses of 35Gy 10 or greater but with an increased incidence of esophagitis. The objective of this planning study was to assess the potential impact of esophageal-sparing IMRT (ES-IMRT) compared to the current standard of care using parallel-opposed pair beams (POP)., Methods: In this study, 15 patients with lung cancer treated to a dose of 30Gy in 10 fractions between August 2015 and January 2016 were identified. Radiation treatment plans were optimized using ES-IMRT by limiting the max esophagus point dose to 24Gy. Using published Lyman-Kutcher-Burman normal tissue complication probabilities (LKB-NTCP) models, both plans were evaluated for the likelihood of esophagitis (≥ grade 2) and pneumonitis (≥ grade 2)., Results: Using ES-IMRT, the median esophageal and lung mean doses reduced from 16 and 8Gy to 7 and 7Gy, respectively. Using the LKB models, the theoretical probability of symptomatic esophagitis and pneumonitis reduced from 13 to 2%, and from 5 to 3%, respectively. The median normalize total dose (NTD mean) accounting for fraction size for the GTV and PTV of the clinically approved POP plans compared to the ES-IMRT plans were similar., Conclusion: Advanced radiotherapy techniques such as ES-IMRT may have clinical utility in reducing treatment-related toxicity in advanced lung cancer patients. Our data suggests that the rate of esophagitis can be reduced without compromising local control.

Objective: The aim of this work was to investigate whether quantitative dual-energy CT (DECT) imaging is feasible for small animal irradiators with an integrated cone-beam CT (CBCT) system., Methods: The optimal imaging protocols were determined by analyzing different energy combinations and dose levels. The influence of beam hardening effects and the performance of a beam hardening correction (BHC) were investigated. In addition, two systems from different manufacturers were compared in terms of errors in the extracted effective atomic numbers (Z eff ) and relative electron densities (ρ e ) for phantom inserts with known elemental compositions and relative electron densities., Results: The optimal energy combination was determined to be 50 and 90 kVp. For this combination, Z eff and ρ e can be extracted with a mean error of 0.11 and 0.010, respectively, at a dose level of 60 cGy., Conclusion: Quantitative DECT imaging is feasible for small animal irradiators with an integrated CBCT system. To obtain the best results, optimizing the imaging protocols is required. Well-separated X-ray spectra and a sufficient dose level should be used to minimize the error and noise for Z eff and ρ e . When no BHC is applied in the image reconstruction, the size of the calibration phantom should match the size of the imaged object to limit the influence of beam hardening effects. No significant differences in Z eff and ρ e errors are observed between the two systems from different manufacturers. Advances in knowledge: This is the first study that investigates quantitative DECT imaging for small animal irradiators with an integrated CBCT system.

Glioblastoma multiforme (GBM) is the most common malignant brain tumor in adults. The current standard of care includes surgery followed by radiotherapy (RT) and chemotherapy with temozolomide (TMZ). Treatment often fails due to the radiation resistance and intrinsic or acquired TMZ resistance of a small percentage of cells with stem cell-like behavior (CSC). The NOTCH signaling pathway is expressed and active in human glioblastoma and NOTCH inhibitors attenuate tumor growth in vivo in xenograft models. Here we show using an image guided micro-CT and precision radiotherapy platform that a combination of the clinically approved NOTCH/γ-secretase inhibitor (GSI) RO4929097 with standard of care (TMZ + RT) reduces tumor growth and prolongs survival compared to dual combinations. We show that GSI in combination with RT and TMZ attenuates proliferation, decreases 3D spheroid growth and results into a marked reduction in clonogenic survival in primary and established glioma cell lines. We found that the glioma stem cell marker CD133, SOX2 and Nestin were reduced following combination treatments and NOTCH inhibitors albeit in a different manner. These findings indicate that NOTCH inhibition combined with standard of care treatment has an anti-glioma stem cell effect which provides an improved survival benefit for GBM and encourages further translational and clinical studies., Competing Interests: The authors have declared that no competing interests exist.

Optical cone-beam computed tomographic (CBCT) scanning of 3D radiochromic dosimeters may provide a practical method for 3D dose verification in radiation therapy. However, in cone-beam geometry stray light contaminates the projection images, degrading the accuracy of reconstructed linear attenuation coefficients. Stray light was measured using a beam pass aperture array (BPA) and structured illumination methods. The stray-to-primary ray ratio (SPR) along the central axis was found to be 0.24 for a 5% gelatin hydrogel, representative of radiochromic hydrogels. The scanner was modified by moving the spectral filter from the detector to the source, changing the light's spatial fluence pattern and lowering the acceptance angle by extending distance between the source and object. These modifications reduced the SPR significantly from 0.24 to 0.06. The accuracy of the reconstructed linear attenuation coefficients for uniform carbon black liquids was compared to independent spectrometer measurements. Reducing the stray light increased the range of accurate transmission readings. In order to evaluate scanner performance for the more challenging application to small field dosimetry, a carbon black finger gel phantom was prepared. Reconstructions of the phantom from CBCT and fan-beam CT scans were compared. The modified source resulted in improved agreement. Subtraction of residual stray light, measured with BPA or structured illumination from each projection further improved agreement. Structured illumination was superior to BPA for measuring stray light for the smaller 1.2 and 0.5 cm diameter phantom fingers. At the costs of doubling the scanner size and tripling the number of scans, CBCT reconstructions of low-scattering hydrogel dosimeters agreed with those of fan-beam CT scans.

Background and Purpose: Glioblastoma multiforme is the most common malignant brain tumor. Standard treatment including surgery, radiotherapy and chemotherapy with temozolomide is not curative. There is a great need for in vitro and in vivo models closely mimicking clinical practice to ensure better translation of novel preclinical findings., Methods and Materials: A 3D spheroid model was established using the U87MG cell line. The efficacy of temozolomide, RT and combinations was assessed using growth delay assays. Orthotopic glioblastoma tumors were established, different radiation doses delivered based on micro-CT based treatment planning (SmART-plan) and dose volume histograms (DVH) were determined. Tumor growth was monitored using bioluminescent imaging., Results: 3D spheroid cultures showed a dose-dependent growth delay upon single and combination treatments. Precise uniform radiation was achieved in all in vivo treatment groups at all doses tested, and DVHs showed accurate dose coverage in the planning target volume which resulted in tumor growth delay., Conclusion: We demonstrate that 3D spheroid technology can be reliably used for treatment efficacy evaluation and that mimicking a clinical setting is also possible in small animals. Both these in vitro and in vivo techniques can be combined for clinically relevant testing of novel drugs combined with radiation., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Advances in precision small animal radiotherapy hardware enable the delivery of increasingly complicated dose distributions on the millimeter scale. Manual creation and evaluation of treatment plans becomes difficult or even infeasible with an increasing number of degrees of freedom for dose delivery and available image data. The goal of this work is to develop an optimisation model that determines beam-on times for a given beam configuration, and to assess the feasibility and benefits of an automated treatment planning system for small animal radiotherapy. The developed model determines a Pareto optimal solution using operator-defined weights for a multiple-objective treatment planning problem. An interactive approach allows the planner to navigate towards, and to select the Pareto optimal treatment plan that yields the most preferred trade-off of the conflicting objectives. This model was evaluated using four small animal cases based on cone-beam computed tomography images. Resulting treatment plan quality was compared to the quality of manually optimised treatment plans using dose-volume histograms and metrics. Results show that the developed framework is well capable of optimising beam-on times for 3D dose distributions and offers several advantages over manual treatment plan optimisation. For all cases but the simple flank tumour case, a similar amount of time was needed for manual and automated beam-on time optimisation. In this time frame, manual optimisation generates a single treatment plan, while the inverse planning system yields a set of Pareto optimal solutions which provides quantitative insight on the sensitivity of conflicting objectives. Treatment planning automation decreases the dependence on operator experience and allows for the use of class solutions for similar treatment scenarios. This can shorten the time required for treatment planning and therefore increase animal throughput. In addition, this can improve treatment standardisation and comparability of research data within studies and among different institutes.

Recently, precision irradiators integrated with a high-resolution CT imaging device became available for pre-clinical studies. These research platforms offer significant advantages over older generations of animal irradiators in terms of precision and accuracy of image-guided radiation targeting. These platforms are expected to play a significant role in defining experiments that will allow translation of research findings to the human clinical setting. In the field of radiotherapy, but also others such as neurology, the platforms create unique opportunities to explore e.g. the synergy between radiation and drugs or other agents. To fully exploit the advantages of this new technology, accurate methods are needed to plan the irradiation and to calculate the three-dimensional radiation dose distribution in the specimen. To this end, dedicated treatment planning systems are needed. In this review we will discuss specific issues for precision irradiation of small animals, we will describe the workflow of animal treatment planning, and we will examine several dose calculation algorithms (factorization, superposition-convolution, Monte Carlo simulation) used for animal irradiation with kilovolt photon beams. Issues such as dose reporting methods, photon scatter, tissue segmentation and motion will also be discussed briefly., (Copyright © 2014. Published by Elsevier GmbH.)

Purpose: In lung cancer radiation therapy, the dose constraints are determined mostly by healthy lung toxicity. Preclinical microirradiators are a new tool to evaluate treatment strategies closer to clinical irradiation devices. In this study, we quantified local changes in lung density symptomatic of radiation-induced lung fibrosis (RILF) after partial lung irradiation in mice by using a precision image-guided small animal irradiator integrated with micro-computed tomography (CT) imaging., Methods and Materials: C57BL/6 adult male mice (n=76) were divided into 6 groups: a control group (0 Gy) and groups irradiated with a single fraction of 4, 8, 12, 16, or 20 Gy using 5-mm circular parallel-opposed fields targeting the upper right lung. A Monte Carlo model of the small animal irradiator was used for dose calculations. Following irradiation, all mice were imaged at regular intervals over 39 weeks (10 time points total). Nonrigid deformation was used to register the initial micro-CT scan to all subsequent scans., Results: Significant differences could be observed between the 3 highest (>10 Gy) and 3 lowest irradiation (<10 Gy) dose levels. A mean difference of 120 ± 10 HU between the 0- and 20-Gy groups was observed at week 39. RILF was found to be spatially limited to the irradiated portion of the lung., Conclusions: The data suggest that the severity of RILF in partial lung irradiation compared to large field irradiation in mice for the same dose is reduced, and therefore higher doses can be tolerated., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Purpose: To compare the dosimetric and geometric properties of a commercial x-ray based image-guided small animal irradiation system, installed at three institutions and to establish a complete and broadly accessible commissioning procedure., Methods: The system consists of a 225 kVp x-ray tube with fixed field size collimators ranging from 1 to 44 mm equivalent diameter. The x-ray tube is mounted opposite a flat-panel imaging detector, on a C-arm gantry with 360° coplanar rotation. Each institution performed a full commissioning of their system, including half-value layer, absolute dosimetry, relative dosimetry (profiles, percent depth dose, and relative output factors), and characterization of the system geometry and mechanical flex of the x-ray tube and detector. Dosimetric measurements were made using Farmer-type ionization chambers, small volume air and liquid ionization chambers, and radiochromic film. The results between the three institutions were compared., Results: At 225 kVp, with 0.3 mm Cu added filtration, the first half value layer ranged from 0.9 to 1.0 mm Cu. The dose-rate in-air for a 40 × 40 mm(2) field size, at a source-to-axis distance of 30 cm, ranged from 3.5 to 3.9 Gy/min between the three institutions. For field sizes between 2.5 mm diameter and 40 × 40 mm(2), the differences between percent depth dose curves up to depths of 3.5 cm were between 1% and 4% on average, with the maximum difference being 7%. The profiles agreed very well for fields >5 mm diameter. The relative output factors differed by up to 6% for fields larger than 10 mm diameter, but differed by up to 49% for fields ≤5 mm diameter. The mechanical characteristics of the system (source-to-axis and source-to-detector distances) were consistent between all three institutions. There were substantial differences in the flex of each system., Conclusions: With the exception of the half-value layer, and mechanical properties, there were significant differences between the dosimetric and geometric properties of the three systems. This underscores the need for careful commissioning of each individual system for use in radiobiological experiments.

Small animal models are crucial to link molecular discoveries and implementation of clinically relevant therapeutics in oncology. Using these models requires noninvasive imaging techniques to monitor disease progression and therapy response. Micro-computed tomography (CT) is less studied for the in vivo monitoring of murine intracranial tumors and traditionally suffers from poor soft tissue contrast, whereas bioluminescence imaging (BLI) is known for its sensitivity but is not frequently employed for quantifying tumor volume. A widely used orthotopic glioblastoma multiforme (GBM) tumor model was applied in nude mice, and tumor growth was evaluated by BLI and contrast-enhanced microCT imaging. A strong correlation was observed between CT volume and BLI-integrated intensity (Pearson coefficient (r)  =  .85, p  =  .0002). Repeated contouring of contrast-enhanced microCT-delineated tumor volumes achieved an intraobserver average pairwise overlap ratio of 0.84 and an average tumor volume coefficient of variance of 0.11. MicroCT-delineated tumor size was found to correlate with tumor size obtained via histologic analysis (Pearson coefficient (r)  =  .88, p  =  .005). We conclude that BLI intensity can be used to derive tumor volume but that the use of both contrast-enhanced microCT and BLI provides complementary tumor growth information, which is particularly useful for modern small animal irradiation devices that make use of microCT and BLI for treatment planning, targeting, and monitoring.

Background and Purpose: Image-guided equipment for precision irradiation of small animals for pre-clinical radiotherapy research became recently available. To enable downscaled radiotherapy studies that can be translated into human radiotherapy knowledge, a treatment planning system for pre-clinical studies is required., Material and Methods: A dedicated treatment planning system (SmART-Plan) for small animal radiotherapy studies was developed. It is based on Monte Carlo simulation of particle transport in an animal. The voxel geometry is derived from the onboard cone beam CT imaging panel. SmART-Plan was validated using radiochromic film (RCF) dosimetry in various phantoms: uniform, multislab and a realistic plasticized mouse geometry., Results: Good agreement was obtained between SmART-Plan dose calculations and RCF dose measurements in all phantoms. For various delivered plans agreement was obtained within 10% for the majority of the targeted dose region, with larger differences between 10% and 20% near the penumbra regions and for the smallest 1mm collimator. Absolute depth and lateral dose distributions showed better agreement for 5 and 15-mm collimators than for a 1-mm collimator, indicating that accurate dose prediction for the smallest field sizes is difficult., Conclusion: SmART-Plan offers a useful dose calculation tool for pre-clinical small animal irradiation studies., (Copyright © 2013 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)

Precision image-guided small animal radiotherapy is rapidly advancing through the use of dedicated micro-irradiation devices. However, precise modeling of these devices in model-based dose-calculation algorithms such as Monte Carlo (MC) simulations continue to present challenges due to a combination of very small beams, low mechanical tolerances on beam collimation, positioning and long calculation times. The specific intent of this investigation is to introduce and demonstrate the viability of a fast analytical source model (AM) for use in either investigating improvements in collimator design or for use in faster dose calculations. MC models using BEAMnrc were developed for circular and square fields sizes from 1 to 25 mm in diameter (or side) that incorporated the intensity distribution of the focal spot modeled after an experimental pinhole image. These MC models were used to generate phase space files (PSFMC) at the exit of the collimators. An AM was developed that included the intensity distribution of the focal spot, a pre-calculated x-ray spectrum, and the collimator-specific entrance and exit apertures. The AM was used to generate photon fluence intensity distributions (ΦAM) and PSFAM containing photons radiating at angles according to the focal spot intensity distribution. MC dose calculations using DOSXYZnrc in a water and mouse phantom differing only by source used (PSFMC versus PSFAM) were found to agree within 7% and 4% for the smallest 1 and 2 mm collimator, respectively, and within 1% for all other field sizes based on depth dose profiles. PSF generation times were approximately 1200 times faster for the smallest beam and 19 times faster for the largest beam. The influence of the focal spot intensity distribution on output and on beam shape was quantified and found to play a significant role in calculated dose distributions. Beam profile differences due to collimator alignment were found in both small and large collimators sensitive to shifts of 1 mm with respect to the central axis.

The effect of oral hyaluronan (HA) on bone loss in ovariectomized (OVX) 3-month-old rats was measured using serum markers of bone turnover and bone mineral density. OVX rats were administered 1 mg/kg HA (OVX + HA) or phosphate-buffered saline (PBS) (OVX + PBS) by oral gavage (5 days/week for 54 days). Additional controls included sham ovariectomy with PBS gavage (Sham + PBS) and no treatment. Oral administration of HA resulted in approximately 50% (p < 0.05) increases in serum HA. Gel filtration analyses showed this was high molecular weight HA (300-500 kDa). Osteopenia was mild due to the young age of the animals. Thus, ovariectomy resulted in a 30% increase in serum collagen N-terminal telopeptides (p < 0.001), a 20% increase in serum nitrate/nitrite levels (p = 0.05), and a 5-6% decrease in femur bone mineral density/content (p < 0.05). HA gavage blunted the development of osteopenia in this model as determined by preventing the 30% increase in serum collagen N-terminal telopeptide levels (p < 0.001) and by reducing bone mineral content loss from 6 to 4%. These results show that oral supplements of HA (gavage solution, 0.12% solution) significantly reduce bone turnover associated with mild osteopenia in rats.

Dual-energy computed tomography (CT) enables 3-dimensional,noninvasive, and nondestructive imaging with material separation. Dual-energy CT is generally used to segment hydrated tissues within the clinical context. We apply dual-energy CT to an ancient Egyptian mummy and present several techniques designed to separate bone from desiccated tissue and resin. Automated and semiautomated dual-energy CT techniques are compared to manual segmentation and thresholding-based techniques. Semiautomated techniques enable substantial reductions in operator time compared to manual segmentation.

A novel approach to read out radiochromic film was introduced recently by the manufacturer of GafChromic film. In this study, the performance of this triple-channel film dosimetry method was compared against the conventional single-red-channel film dosimetry procedure, with and without inclusion of a pre-irradiation (pre-IR) film scan, using EBT2 film and kilo- and megavoltage photon beams up to 10 Gy. When considering regions of interest averaged doses, the triple-channel method and both single-channel methods produced equivalent results. Absolute dose discrepancies between the triple-channel method, both single-channel methods and the treatment planning system calculated dose values, were no larger than 5 cGy for dose levels up to 2.2 Gy. Signal to noise in triple-channel dose images was found to be similar to signal to noise in single-channel dose images. The accuracy of resulting dose images from the triple- and single-channel methods with inclusion of pre-IR film scan was found to be similar. Results of a comparison of EBT2 data from a kilovoltage depth dose experiment to corresponding Monte Carlo depth dose data produced dose discrepancies of 9.5 ± 12 cGy and 7.6 ± 6 cGy for the single-channel method with inclusion of a pre-IR film scan and the triple-channel method, respectively. EBT2 showed to be energy sensitive at low kilovoltage energies with response differences of 11.9% and 15.6% in the red channel at 2 Gy between 50-225 kVp and 80-225 kVp photon spectra, respectively. We observed that the triple-channel method resulted in non-uniformity corrections of ±1% and consistency values of 0-3 cGy for the batches and dose levels studied. Results of this study indicate that the triple-channel radiochromic film read-out method performs at least as well as the single-channel method with inclusion of a pre-IR film scan, reduces film non-uniformity and saves time with elimination of a pre-IR film scan.

Purpose: Novel small animal precision microirradiators (micro-IR) are becoming available for preclinical use and are often equipped with onboard imaging (OBI) devices. We investigated the use of OBI as a means to infer the accuracy of the delivered treatment plan., Methods: Monte Carlo modeling of the micro-IR including an elliptical Gaussian electron beam incident on the x-ray tube was used to score dose and to continue photon transport to the plane of the OBI device. A model of the OBI detector response was used to generate simulated onboard images. Experimental OBI was performed at 225 kVp, gain∕offset and scatter-glare were corrected. Simulated and experimentally obtained onboard images of phantoms and a mouse specimen were compared for a range of photon beam sizes from 2.5 cm down to 0.1 cm., Results: Simulated OBI can be used in small animal radiotherapy to determine if a treatment plan was delivered according to the prescription within an uncertainty of 5% for beams as small as 4 mm in diameter. For collimated beams smaller than 4 mm, beam profile differences remain primarily in the penumbra region of the smallest beams, which may be tolerable for specific preclinical micro-IR investigations., Conclusions: Comparing simulated to acquired OBI during small animal treatment radiotherapy represents a useful treatment delivery tool.

This work compares Monte Carlo (MC) dose calculations for (125)I and (103)Pd low-dose rate (LDR) brachytherapy sources performed in virtual phantoms containing a series of human soft tissues of interest for brachytherapy. The geometries are segmented (tissue type and density assignment) based on simulated single energy computed tomography (SECT) and dual energy (DECT) images, as well as the all-water TG-43 approach. Accuracy is evaluated by comparison to a reference MC dose calculation performed in the same phantoms, where each voxel's material properties are assigned with exactly known values. The objective is to assess potential dose calculation accuracy gains from DECT. A CT imaging simulation package, ImaSim, is used to generate CT images of calibration and dose calculation phantoms at 80, 120, and 140 kVp. From the high and low energy images electron density ρ(e) and atomic number Z are obtained using a DECT algorithm. Following a correction derived from scans of the calibration phantom, accuracy on Z and ρ(e) of ±1% is obtained for all soft tissues with atomic number Z ∊ [6,8] except lung. GEANT4 MC dose calculations based on DECT segmentation agreed with the reference within ±4% for (103)Pd, the most sensitive source to tissue misassignments. SECT segmentation with three tissue bins as well as the TG-43 approach showed inferior accuracy with errors of up to 20%. Using seven tissue bins in our SECT segmentation brought errors within ±10% for (103)Pd. In general (125)I dose calculations showed higher accuracy than (103)Pd. Simulated image noise was found to decrease DECT accuracy by 3-4%. Our findings suggest that DECT-based segmentation yields improved accuracy when compared to SECT segmentation with seven tissue bins in LDR brachytherapy dose calculation for the specific case of our non-anthropomorphic phantom. The validity of our conclusions for clinical geometry as well as the importance of image noise in the tissue segmentation procedure deserves further experimental investigation.

Background and Purpose: Dual energy CT (DECT) imaging can provide both the electron density ρ(e) and effective atomic number Z(eff), thus facilitating tissue type identification. This paper investigates the accuracy of a dual source DECT scanner by means of measurements and simulations. Previous simulation work suggested improved Monte Carlo dose calculation accuracy when compared to single energy CT for low energy photon brachytherapy, but lacked validation. As such, we aim to validate our DECT simulation model in this work., Materials and Methods: A cylindrical phantom containing tissue mimicking inserts was scanned with a second generation dual source scanner (SOMATOM Definition FLASH) to obtain Z(eff) and ρ(e). A model of the scanner was designed in ImaSim, a CT simulation program, and was used to simulate the experiment., Results: Accuracy of measured Z(eff) (labelled Z) was found to vary from -10% to 10% from low to high Z tissue substitutes while the accuracy on ρ(e) from DECT was about 2.5%. Our simulation reproduced the experiments within ±5% for both Z and ρ(e)., Conclusions: A clinical DECT scanner was able to extract Z and ρ(e) of tissue substitutes. Our simulation tool replicates the experiments within a reasonable accuracy., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Precise, noninvasive analysis and quantification of in vivo body composition is essential for research involving longitudinal, small-animal disease models. We investigated the feasibility and precision of a rapid, flat-panel μCT scanner to report whole body adipose tissue volume (ATV), lean tissue volume (LTV), skeletal tissue volume (STV), and bone mineral content (BMC) in 25 postmortem female and 52 live male Sprague-Dawley rats. μCT images, acquired in three 90-mm segments and reconstructed with 308 μm of isotropic voxel spacing, formed contiguous image volumes of each entire rat specimen. Three signal-intensity thresholds (determined to be -186, 5, and 155 HU) were used to classify each voxel as adipose, lean, or skeletal tissue, respectively. Tissue masses from the volume fractions of ATV, LTV, and STV were calculated from assumed tissue densities of 0.95, 1.05, and 1.92 g/cm(-3), respectively. A CT-derived total mass was calculated for each rat and compared with the gravimetrically measured mass, which differed on average for the postmortem female and the live male group by 2.5 and 1.1%, respectively. To evaluate the accuracy of the CT-derived body composition technique, following the live male study excised muscle tissue in the lower right leg of all rats in group B were compared with the image-derived LT measurement of the same regional compartment and found to differ on average by 2.2%. Through repeated CT measurements of postmortem specimens, the whole body ATV, LTV, STV, and BMC measurement analysis gave a precision value of ±0.6, 1.9, 1.7, and 0.5% of the average value, respectively.

Micro-CT has become a powerful tool for small animal research, having the ability to obtain high-resolution in vivo and ex vivo images for analyzing bone mineral content, organ vasculature, and bone microarchitecture extraction. The use of exogenous contrast agents further extends the use of micro-CT techniques, but despite advancements in contrast agents, single-energy micro-CT is still limited in cases where two different materials share similar grey-scale intensity values. This study specifically addresses the development of multiple-energy cone-beam micro-CT, for applications where bone must be separated from blood vessels filled with a Pb-based contrast material (Microfil) in ex vivo studies of rodents and tissue specimens. The authors report the implementation of dual- and triple-energy CT algorithms for material-specific imaging using postreconstruction decomposition of micro-CT data; the algorithms were implemented on a volumetric cone-beam micro-CT scanner (GE Locus Ultra). For the dual-energy approach, extrinsic filtration was applied to the x-ray beam to produce spectra with different proportions of x rays above the K edge of Pb. The optimum x-ray tube energies (140 kVp filtered with 1.45 mm Cu and 96 kVp filtered with 0.3 mm Pb) that maximize the contrast between bone and Microfil were determined through numerical simulation. For the triple-energy decomposition, an additional low-energy spectrum (70 kVp, no added filtration) was used. The accuracy of decomposition was evaluated through simulations and experimental verification of a phantom containing a cortical bone simulating material (SB3), Microfil, and acrylic. Using simulations and phantom experiments, an accuracy greater than 95% was achieved in decompositions of bone and Microfil (for noise levels lower than 11 HU), while soft tissue was separated with accuracy better than 99%. The triple-energy technique demonstrated a slightly higher, but not significantly different, decomposition accuracy than the dual-energy technique for the same achieved noise level in the micro-CT images acquired at the multiple energies. The dual-energy technique was applied to the decomposition of an ex vivo rat specimen perfused with Microfil; successful decomposition of the bone and Microfil was achieved, enabling the visualization and characterization of the vasculature both in areas where the vessels traverse soft tissue and when they are surrounded by bone. In comparison, in single energy micro-CT, vessels surrounded by bone could not be distinguished from the cortical bone, based on grey-scale intensity alone. This work represents the first postreconstruction application of material-specific decomposition that directly takes advantage of the K edge characteristics of a contrast material injected into an animal specimen; the application of the technique resulted in automatic, accurate segmentation of 3D micro-CT images into bone, vessel, and tissue components. The algorithm uses only reconstructed images, rather than projection data, and is calibrated by an operator with signal values in regions identified as being comprised entirely of either cortical bone, contrast-enhanced vessel, or soft tissue; these required calibration values are observed directly within reconstructed CT images acquired at the multiple energies. These features facilitate future implementation on existing research micro-CT systems.