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Title: SU-F-J-220: Micro-CT Based Quantification of Mouse Brain Vasculature: The Effects of Acquisition Technique and Contrast Material

Abstract

Purpose: Cognitive impairment from radiation therapy to the brain may be linked to the loss of total blood volume in the brain. To account for brain injury, it is crucial to develop an understanding of blood volume loss as a result of radiation therapy. This study investigates µCT based quantification of mouse brain vasculature, focusing on the effect of acquisition technique and contrast material. Methods: Four mice were scanned on a µCT scanner (Siemens Inveon). The reconstructed voxel size was 18µm3 and all protocols were Hounsfield Unit (HU) calibrated. The mice were injected with 40mg of gold nanoparticles (MediLumine) or 100µl of Exitron 12000 (Miltenyi Biotec). Two acquisition techniques were also performed. A single kVp technique scanned the mouse once using an x-ray beam of 80kVp and segmentation was completed based on a threshold of HU values. The dual kVp technique scanned the mouse twice using 50kVp and 80kVp, this segmentation was based on the ratio of the HU value of the two kVps. After image reconstruction and segmentation, the brain blood volume was determined as a percentage of the total brain volume. Results: For the single kVp acquisition at 80kVp, the brain blood volume had an average of 3.5%more » for gold and 4.0% for Exitron 12000. Also at 80kVp, the contrast-noise ratio was significantly better for images acquired with the gold nanoparticles (2.0) than for those acquired with the Exitron 12000 (1.4). The dual kVp acquisition shows improved separation of skull from vasculature, but increased image noise. Conclusion: In summary, the effects of acquisition technique and contrast material for quantification of mouse brain vasculature showed that gold nanoparticles produced more consistent segmentation of brain vasculature than Exitron 12000. Also, dual kVp acquisition may improve the accuracy of brain vasculature quantification, although the effect of noise amplification warrants further study.« less

Authors:
; ; ; ;  [1]
  1. University of Cincinnati College of Medicine, Cincinnati, OH (United States)
Publication Date:
OSTI Identifier:
22642247
Resource Type:
Journal Article
Resource Relation:
Journal Name: Medical Physics; Journal Volume: 43; Journal Issue: 6; Other Information: (c) 2016 American Association of Physicists in Medicine; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 61 RADIATION PROTECTION AND DOSIMETRY; ACCURACY; BLOOD; BRAIN; COMPUTERIZED TOMOGRAPHY; IMAGE PROCESSING; IMAGES; INJURIES; MICE; NANOPARTICLES; RADIOTHERAPY; SKULL

Citation Formats

Tipton, C, Lamba, M, Qi, Z, LaSance, K, and Tipton, C. SU-F-J-220: Micro-CT Based Quantification of Mouse Brain Vasculature: The Effects of Acquisition Technique and Contrast Material. United States: N. p., 2016. Web. doi:10.1118/1.4956128.
Tipton, C, Lamba, M, Qi, Z, LaSance, K, & Tipton, C. SU-F-J-220: Micro-CT Based Quantification of Mouse Brain Vasculature: The Effects of Acquisition Technique and Contrast Material. United States. doi:10.1118/1.4956128.
Tipton, C, Lamba, M, Qi, Z, LaSance, K, and Tipton, C. 2016. "SU-F-J-220: Micro-CT Based Quantification of Mouse Brain Vasculature: The Effects of Acquisition Technique and Contrast Material". United States. doi:10.1118/1.4956128.
@article{osti_22642247,
title = {SU-F-J-220: Micro-CT Based Quantification of Mouse Brain Vasculature: The Effects of Acquisition Technique and Contrast Material},
author = {Tipton, C and Lamba, M and Qi, Z and LaSance, K and Tipton, C},
abstractNote = {Purpose: Cognitive impairment from radiation therapy to the brain may be linked to the loss of total blood volume in the brain. To account for brain injury, it is crucial to develop an understanding of blood volume loss as a result of radiation therapy. This study investigates µCT based quantification of mouse brain vasculature, focusing on the effect of acquisition technique and contrast material. Methods: Four mice were scanned on a µCT scanner (Siemens Inveon). The reconstructed voxel size was 18µm3 and all protocols were Hounsfield Unit (HU) calibrated. The mice were injected with 40mg of gold nanoparticles (MediLumine) or 100µl of Exitron 12000 (Miltenyi Biotec). Two acquisition techniques were also performed. A single kVp technique scanned the mouse once using an x-ray beam of 80kVp and segmentation was completed based on a threshold of HU values. The dual kVp technique scanned the mouse twice using 50kVp and 80kVp, this segmentation was based on the ratio of the HU value of the two kVps. After image reconstruction and segmentation, the brain blood volume was determined as a percentage of the total brain volume. Results: For the single kVp acquisition at 80kVp, the brain blood volume had an average of 3.5% for gold and 4.0% for Exitron 12000. Also at 80kVp, the contrast-noise ratio was significantly better for images acquired with the gold nanoparticles (2.0) than for those acquired with the Exitron 12000 (1.4). The dual kVp acquisition shows improved separation of skull from vasculature, but increased image noise. Conclusion: In summary, the effects of acquisition technique and contrast material for quantification of mouse brain vasculature showed that gold nanoparticles produced more consistent segmentation of brain vasculature than Exitron 12000. Also, dual kVp acquisition may improve the accuracy of brain vasculature quantification, although the effect of noise amplification warrants further study.},
doi = {10.1118/1.4956128},
journal = {Medical Physics},
number = 6,
volume = 43,
place = {United States},
year = 2016,
month = 6
}
  • Angiogenesis is widely investigated in conjunction with cancer development, in particular because of the possibility of early stage detection and of new therapeutic strategies. However, such studies are negatively affected by the limitations of imaging techniques in the detection of microscopic blood vessels (diameter 3-5 {micro}m) grown under angiogenic stress. We report that synchrotron-based X-ray imaging techniques with very high spatial resolution can overcome this obstacle, provided that suitable contrast agents are used. We tested different contrast agents based on gold nanoparticles (AuNPs) for the detection of cancer-related angiogenesis by synchrotron microradiology, microtomography and high resolution X-ray microscopy. Among themmore » only bare-AuNPs in conjunction with heparin injection provided sufficient contrast to allow in vivo detection of small capillary species (the smallest measured lumen diameters were 3-5 {micro}m). The detected vessel density was 3-7 times higher than with other nanoparticles. We also found that bare-AuNPs with heparin allows detecting symptoms of local extravascular nanoparticle diffusion in tumor areas where capillary leakage appeared. Although high-Z AuNPs are natural candidates as radiology contrast agents, their success is not guaranteed, in particular when targeting very small blood vessels in tumor-related angiography. We found that AuNPs injected with heparin produced the contrast level needed to reveal--for the first time by X-ray imaging--tumor microvessels with 3-5 {micro}m diameter as well as extravascular diffusion due to basal membrane defenestration. These results open the interesting possibility of functional imaging of the tumor microvasculature, of its development and organization, as well as of the effects of anti-angiogenic drugs.« less
  • Purpose: Radiation therapy of CNS tumors damages the blood-brain barrier (BBB) and normal brain tissue. Our aims were to characterize the short- and long-term effects of fractionated radiotherapy (FRT) on cerebral microvasculature in mice and to investigate the mechanism of change in BBB permeability in mice. Methods and Materials: Intravital microscopy and a cranial window technique were used to measure BBB permeability to fluorescein isothiocyanate (FITC)-dextran and leukocyte endothelial interactions before and after cranial irradiation. Daily doses of 2 Gy were delivered 5 days/week (total, 40 Gy). We immunostained the molecules to detect the expression of glial fibrillary acidic proteinmore » and to demonstrate astrocyte activity in brain parenchyma. To relate the permeability changes to endothelial ultrastructural changes, we used electron microscopy. Results: Blood-brain barrier permeability did not increase significantly until 90 days after FRT, at which point it increased continuously until 180 days post-FRT. The number of adherent leukocytes did not increase during the study. The number of astrocytes in the cerebral cortex increased significantly; vesicular activity in endothelial cells increased beginning 90 days after irradiation, and most tight junctions stayed intact, although some were shorter and less dense at 120 and 180 days. Conclusions: The cellular and microvasculature response of the brain to FRT is mediated through astrogliosis and ultrastructural changes, accompanied by an increase in BBB permeability. The response to FRT is delayed as compared with single-dose irradiation treatment, and does not involve leukocyte adhesion. However, FRT induces an increase in the BBB permeability, as in the case of single-dose irradiation.« less
  • Purpose: Damage to normal tissue is a limiting factor in clinical radiotherapy (RT). We tested the hypothesis that the presence of tumor alters the response of normal tissues to irradiation using a rat in situ brain tumor model. Methods and Materials: Intravital microscopy was used with a rat cranial window to assess the in situ effect of rat C6 glioma on peritumoral tissue with and without RT. The RT regimen included 40 Gy at 8 Gy/day starting Day 5 after tumor implant. Endpoints included blood-brain barrier permeability, clearance index, leukocyte-endothelial interactions and staining for vascular endothelial growth factor (VEGF) glialmore » fibrillary acidic protein, and apoptosis. To characterize the system response to RT, animal survival and tumor surface area and volume were measured. Sham experiments were performed on similar animals implanted with basement membrane matrix absent of tumor cells. Results: The presence of tumor alone increases permeability but has little effect on leukocyte-endothelial interactions and astrogliosis. Radiation alone increases tissue permeability, leukocyte-endothelial interactions, and astrogliosis. The highest levels of permeability and cell adhesion were seen in the model that combined tumor and irradiation; however, the presence of tumor appeared to reduce the volume of rolling leukocytes. Unirradiated tumor and peritumoral tissue had poor clearance. Irradiated tumor and peritumoral tissue had a similar clearance index to irradiated and unirradiated sham-implanted animals. Radiation reduces the presence of VEGF in peritumoral normal tissues but did not affect the amount of apoptosis in the normal tissue. Apoptosis was identified in the tumor tissue with and without radiation. Conclusions: We developed a novel approach to demonstrate that the presence of the tumor in a rat intracranial model alters the response of normal tissues to irradiation.« less
  • Purpose: To examine the addition of genetic or pharmacologic inhibition of hypoxia-inducible factor 1α (HIF-1α) to radiation therapy (RT) and vascular endothelial growth factor A (VEGF-A) inhibition (ie trimodality therapy) for soft-tissue sarcoma. Methods and Materials: Hypoxia-inducible factor 1α was inhibited using short hairpin RNA or low metronomic doses of doxorubicin, which blocks HIF-1α binding to DNA. Trimodality therapy was examined in a mouse xenograft model and a genetically engineered mouse model of sarcoma, as well as in vitro in tumor endothelial cells (ECs) and 4 sarcoma cell lines. Results: In both mouse models, any monotherapy or bimodality therapy resulted in tumormore » growth beyond 250 mm{sup 3} within the 12-day treatment period, but trimodality therapy with RT, VEGF-A inhibition, and HIF-1α inhibition kept tumors at <250 mm{sup 3} for up to 30 days. Trimodality therapy on tumors reduced HIF-1α activity as measured by expression of nuclear HIF-1α by 87% to 95% compared with RT alone, and cytoplasmic carbonic anhydrase 9 by 79% to 82%. Trimodality therapy also increased EC-specific apoptosis 2- to 4-fold more than RT alone and reduced microvessel density by 75% to 82%. When tumor ECs were treated in vitro with trimodality therapy under hypoxia, there were significant decreases in proliferation and colony formation and increases in DNA damage (as measured by Comet assay and γH2AX expression) and apoptosis (as measured by cleaved caspase 3 expression). Trimodality therapy had much less pronounced effects when 4 sarcoma cell lines were examined in these same assays. Conclusions: Inhibition of HIF-1α is highly effective when combined with RT and VEGF-A inhibition in blocking sarcoma growth by maximizing DNA damage and apoptosis in tumor ECs, leading to loss of tumor vasculature.« less
  • Purpose: To find effective setting methods to mitigate the irradiation injure in synchrotron radiation microangiography(SRA) by Monte Carlo simulation. Methods: A mouse 1-D head model and a segmented voxel mouse head phantom were simulated by EGSnrc/Dosxyznrc code to investigate the dose enhancement effect of the iodine contrast agent irradiated by a monochromatic synchrotron radiation(SR) source. The influence of, like iodine concentration (IC), vessel width and depth, with and without skull layer protection and the various incident X ray energies, were simulated. The dose enhancement effect and the absolute dose based on the segmented voxel mouse head phantom were evaluated. Results:more » The dose enhancement ratio depends little on the irradiation depth, but strongly on the IC, which is linearly increases with IC. The skull layer protection cannot be ignored in SRA, the 700µm thick skull could decrease 10% of the dose. The incident X-ray energy can significantly affact the dose. E.g. compared to the dose of 33.2keV for 50mgI/ml, the 32.7keV dose decreases 38%, whereas the dose of 33.7 keV increases 69.2%, and the variation will strengthen more with enhanced IC. The segmented voxel mouse head phantom also showed that the average dose enhancement effect and the maximal voxel dose per photon depends little on the iodine voxel volume ratio, but strongly on IC. Conclusion: To decrease dose damage in SRA, the high-Z contrast agent should be used as little as possible, and try to avoid radiating locally the injected position immediately after the contrast agent injection. The fragile vessel containing iodine should avoid closely irradiating. Avoiding irradiating through the no or thin skull region, or appending thin equivalent material from outside to protect is also a better method. As long as SRA image quality is ensured, using incident X-ray energy as low as possible.« less