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3D imaging of transition metals in the zebrafish embryo by x-ray fluorescence microtomography

METALLOMICS 2014年第9期6卷 1648-1655页

作者： Bourassa, Daisy Gleber, Sophie-Charlotte Vogt, Stefan Yi, Hong Will, Fabian Richter, Heiko Shin, Chong Hyun Fahrni, Christoph J. Georgia Inst Technol Sch Chem & Biochem Atlanta GA 30332 USA Georgia Inst Technol Petit Inst Bioengn & Biosci Atlanta GA 30332 USA Argonne Natl Lab Xray Sci Div Adv Photon Source Argonne IL 60439 USA Emory Univ Robert P Apkarian Integrated Electron Microscopy Atlanta GA 30322 USA LLS Rowiak LaserLabSolut GmbH D-30419 Hannover Germany Georgia Inst Technol Sch Biol Atlanta GA 30332 USA

Synchrotron x-ray fluorescence (SxRF) microtomography has emerged as a powerful technique for the 3D visualization of the elemental distribution in biological samples. The mechanical stability, both of the instrument and the specimen, is paramount when acquiring tomographic projection series. By combining the progressive lowering of temperature method (PLT) with femtosecond laser sectioning, we were able to embed, excise, and preserve a zebrafish embryo at 24 hours post fertilization in an x-ray compatible, transparent resin for tomographic elemental imaging. Based on a data set comprised of 60 projections, acquired with a step size of 2 mu m during 100 hours of beam time, we reconstructed the 3D distribution of zinc, iron, and copper using the iterative maximum likelihood expectation maximization (MLEM) reconstruction algorithm. The volumetric elemental maps, which entail over 124 million individual voxels for each transition metal, revealed distinct elemental distributions that could be correlated with characteristic anatomical features at this stage of embryonic development.

关键词： Animals Embryo, Nonmammalian/metabolism Fluorescence Fluorescence Imaging, Three-Dimensional/methods Lasers Time Factors Transition Elements/metabolism x-ray microtomography/methods Zebrafish/embryology

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Optical Tomography of MMP Activity Allows a Sensitive Noninvasive Characterization of the Invasiveness and Angiogenesis of SCC xenografts

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NEOPLASIA 2014年第3期16卷 235-246页

作者： Al Rawashdeh, Wa'el Arns, Susanne Gremse, Felix Ehling, Josef Knuechel-Clarke, Ruth Kray, Stefan Spoeler, Felix Kiessling, Fabian Lederle, Wiltrud Rheinisch Westfael Tech Hsch Aachen Univ Dept Expt Mol Imaging Aachen Germany Rhein Westfal TH Aachen Univ Hosp Aachen Inst Pathol Aachen Germany Rhein Westfal TH Aachen Inst Semicond Elect Aachen Germany

For improved tumor staging and therapy control, imaging biomarkers are of great interest allowing a noninvasive characterization of invasiveness. In squamous epithelial skin and cervix lesions, transition to invasive stages is associated with enhanced matrix metalloproteinase (MMP) activity, increased angiogenesis, and worsened prognosis. Thus, we investigated MMP activity as imaging biomarker of invasiveness and the potential of optical tomography in characterizing the angiogenic and invasive behavior of skin squamous cell carcinoma (SCC) xenografts. MMP activity was measured in vivo in HaCaT-ras A-5RT3 tumors at different angiogenic and invasive stages (onset of angiogenesis, intermediate and highly angiogenic, invasive stage) and after 1 week of sunitinib treatment by fluorescence molecular tomography-microcomputed tomography imaging using an activatable probe. Treatment response was additionally assessed morphologically by optical coherence tomography (OCT). In vivo MMP activity significantly differed between the groups, revealing highest levels in the highly angiogenic, invasive tumors that were confirmed by immunohistochemistry. At the onset of angiogenesis with lowest MMP activity, fibroblasts were detected in the MMP-positive areas, whereas macrophages were absent. Accumulation of both cell types occurred in both invasive groups, again to a significantly higher degree at the most invasive and angiogenic stage. Sunitinib treatment significantly reduced the MMP activity and accumulation of fibroblasts and macrophages and blocked tumor invasion that was additionally visualized by OCT. Human cervical SCCs also showed high MMP activity and a similar stromal composition as the HaCaT xenografts, whereas normal tissue was negative. This study strongly suggests MMP activity as imaging biomarker and demonstrates the high sensitivity of optical tomography in determining tumor invasiveness that can morphologically be supported by OCT.

关键词： Angiogenesis Inhibitors/pharmacology Animals Biomarkers, Tumor/analysis Biomarkers, Tumor/metabolism Carcinoma, Squamous Cell/blood supply Carcinoma, Squamous Cell/drug therapy Carcinoma, Squamous Cell/enzymology Carcinoma, Squamous Cell/pathology Female Humans Indoles/pharmacology Matrix Metalloproteinases/analysis Matrix Metalloproteinases/metabolism Mice, Nude Neovascularization, Pathologic/drug therapy Neovascularization, Pathologic/enzymology Neovascularization, Pathologic/pathology Pyrroles/pharmacology Skin Neoplasms/blood supply Skin Neoplasms/drug therapy Skin Neoplasms/enzymology Skin Neoplasms/pathology Stromal Cells/enzymology Stromal Cells/pathology Tomography, Optical Coherence/methods Tomography, Optical Coherence/methods Uterine Cervical Neoplasms/enzymology Uterine Cervical Neoplasms/pathology xenograft Model Antitumor Assays x-ray microtomography/methods

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Alignment of Biological Apatite Crystallites at First Molar in Human Mandible Cortical Bone

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CRANIO-THE JOURNAL OF CRANIOMANDIBULAR & SLEEP PRACTICE 2012年第1期30卷 32-40页

作者： Morioka, Toshiyuki Matsunaga, Satoru Yoshinari, Masao Ide, Yoshinobu Nakano, Takayoshi Sekine, Hideshi Yajima, Yasutomo Tokyo Dent Coll Oral Hlth Sci Ctr Div Oral Implants Res Mihama Ku 1-2-2 Masago Chiba 2618502 Japan

The aim of this study was to quantitatively clarify the c-axis alignment of biological apatite (BAp) crystallites (hereafter referred to as BAp alignment) in the cortical bone of the human mandible first molar. Six mandible specimens were collected from the cadavers of six dentulous Japanese adults (mean age, 63.0 +/- 12.1 years) held at the Department of Anatomy, Tokyo Dental College. A microbeam x-ray diffraction system was used to determine BAp alignment in the mesiodistal direction. Bone mineral density (BMD) was also measured using 3-dimensional trabecular structure measurement software. The results showed that the degree of BAp alignment in the mesiodistal direction was low in the alveolar area and high at the base of the mandible, suggesting that BAp alignment in the alveolar area is affected by occlusal force. Moreover, it was observed that the correlation between BAp alignment and BMD was small, indicating that BAp alignment and BMD could be independent factors. Therefore, determining BAp alignment was important in the evaluation of bone quality, including bone strength.

关键词： Aged Aged, 80 and over Alveolar Process/ultrastructure Apatites/chemistry Apatites/chemistry Bite Force Bone Density/physiology Cadaver Crystallography Female Humans Image Processing, Computer-Assisted/methods Imaging, Three-Dimensional/methods Male Mandible/ultrastructure Middle Aged Molar Stress, Mechanical x-ray Diffraction/methods x-ray microtomography/methods

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Evaluation of the Palatal Bone for Placement of Orthodontic Mini-Implants in Japanese Adults

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CRANIO-THE JOURNAL OF CRANIOMANDIBULAR & SLEEP PRACTICE 2012年第1期30卷 72-79页

作者： Nakahara, Ken Matsunaga, Satoru Abe, Shinichi Tamatsu, Yuichi Kageyama, Ikuo Hashimoto, Masatsugu Ide, Yoshinobu Tokyo Dent Coll Dept Anat Mihama Ku 1-2-2 Masago Chiba 2618502 Japan

Mini-implants are increasingly being used for orthodontic anchorage in the palate. The anatomical structure of the jaw must be properly evaluated prior to use;however, there are a few research reports providing basic data regarding the palate. Bone thickness was measured and bone morphology evaluated in the palates of Japanese people. The palates of five Japanese adult cadavers and 15 skulls were examined. The samples were imaged and measured using the micro-CT system. In the mid-palatine suture region, the cortical bone had a complex mesh-like structure and was thicker than surrounding areas. Cortical bone thickness varied depending on the site. The mid-palatine suture region is an ideal site for mini-implant insertion;however, because bone and cortical bone thickness markedly decrease in the lateral region, careful attention should be paid when inserting mini-implants in the mid-palatine suture.

关键词： Adolescent Adult Aged Cadaver Cephalometry/methods Cranial Sutures/anatomy & histology Dental Implants Humans Image Processing, Computer-Assisted/methods Imaging, Three-Dimensional/methods Japan Japan Middle Aged Orthodontic Anchorage Procedures/instrumentation Palate, Hard/anatomy & histology x-ray microtomography/methods Young Adult

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Mice Lacking the Calcineurin Inhibitor Rcan2 Have an Isolated Defect of Osteoblast Function

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ENDOCRINOLOGY 2012年第7期153卷 3537-3548页

作者： Bassett, J. H. Duncan Logan, John G. Boyde, Alan Cheung, Moira S. Evans, Holly Croucher, Peter Sun, xiao-yang xu, Sai Murata, Yoshiharu Williams, Graham R. Univ London Imperial Coll Sci Technol & Med Mol Endocrinol Grp Dept Med London W12 0NN England Univ London Queen Mary Coll Ctr Oral Growth & Dev London E1 4NS England Univ Sheffield Mellanby Ctr Bone Res Dept Human Metab Sheffield S10 2RX S Yorkshire England Garvan Inst Med Res Sydney NSW 2010 Australia Nagoya Univ Dept Genet Environm Med Res Inst Nagoya Aichi 4648601 Japan

Calcineurin-nuclear factor of activated T cells signaling controls the differentiation and function of osteoclasts and osteoblasts, and regulator of calcineurin-2 (Rcan2) is a physiological inhibitor of this pathway. Rcan2 expression is regulated by T-3, which also has a central role in skeletal development and bone turnover. To investigate the role of Rcan2 in bone development and maintenance, we characterized Rcan2(-/-) mice and determined its skeletal expression in T-3 receptor (TR) knockout and thyroid-manipulated mice. Rcan2(-/-) mice had normal linear growth but displayed delayed intramembranous ossification, impaired cortical bone formation, and reduced bone mineral accrual during development as well as increased mineralization of adult bone. These abnormalities resulted from an isolated defect in osteoblast function and are similar to skeletal phenotypes of mice lacking the type 2 deiodinase thyroid hormone activating enzyme or with dominant-negative mutations of TR beta, the predominant TR isoform in bone. Rcan2 mRNA was expressed in primary osteoclasts and osteoblasts, and its expression in bone was differentially regulated in TR alpha and TR beta knockout and thyroid-manipulated mice. However, in primary osteoblast cultures, T-3 treatment did not affect Rcan2 mRNA expression or nuclear factor of activated T cells c1 expression and phosphorylation. Overall, these studies establish that Rcan2 regulates osteoblast function and its expression in bone is regulated by thyroid status in vivo. (Endocrinology 153: 3537-3548, 2012)

关键词： Animals Biomechanics Bone and Bones/metabolism Bone Resorption Calcineurin/antagonists & inhibitors Genes, Dominant Mice Mice, Knockout Mice, Knockout Mice, Transgenic Mice, Transgenic Models, Biological Mutation Osteoblasts/cytology Osteoblasts/physiology Phosphorylation Protein Isoforms Proteins/genetics Proteins/physiology RNA, Messenger/metabolism Thyroid Hormone Receptors alpha/metabolism Thyroid Hormone Receptors beta/metabolism T-Lymphocytes/cytology x-ray microtomography/methods

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Optical advances in skeletal imaging applied to bone metastases

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BONE 2011年第1期48卷 106-114页

作者： Snoeks, T. J. A. Khmelinskii, A. Lelieveldt, B. P. F. Kaijzel, E. L. Lowik, C. W. G. M. Leiden Univ Med Ctr Dept Endocrinol NL-2333 ZA Leiden Netherlands Leiden Univ Med Ctr Dept Image Proc LKEB NL-2333 ZA Leiden Netherlands

Optical Imaging has evolved into one of the standard molecular imaging modalities used in pre-clinical cancer research. Bone research however, strongly depends on other imaging modalities such as SPECT, PET, x-ray and mu CT. Each imaging modality has its own specific strengths and weaknesses concerning spatial resolution, sensitivity and the possibility to quantify the signal. An increasing number of bone specific optical imaging models and probes have been developed over the past years. This review gives an overview of optical imaging modalities, models and probes that can be used to study skeletal complications of cancer in small laboratory animals. (C) 2010 Elsevier Inc. All rights reserved.

关键词： Animals Bone Neoplasms/radionuclide imaging Bone Neoplasms/secondary Diagnostic Imaging/methods Forecasting Optics and Photonics/trends Positron-Emission Tomography/methods Tomography, Emission-Computed, Single-Photon/methods x-ray microtomography/methods x-rays x-rays

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What Lies Beneath: In Vivo Stem Cell Imaging

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BIOTECHNIQUES 2011年第4期50卷 223-+页

作者： Perkel, Jeffrey M.

No other branch of science is quite so laden with expectation. But while corralling stem cells for viable therapies, researchers have found themselves asking an entirely separate question: How do we track them?

关键词： Animals Cell Tracking/methods Humans Lung Neoplasms Magnetic Resonance Imaging/methods Magnetite Nanoparticles/chemistry Mesenchymal Stromal Cells/chemistry Mesenchymal Stromal Cells/cytology Oryzias Rats x-ray microtomography/methods Zebrafish

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Three-Dimensional Analysis of the Internal Structure of the Mandibular Condyle in Dentulous and Edentulous Jaws Using Micro-CT

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CRANIO-THE JOURNAL OF CRANIOMANDIBULAR & SLEEP PRACTICE 2009年第2期27卷 78-87页

作者： Sugisaki, Masanori Agematsu, Hiroko Matsunaga, Satoru Saka, Hideki Sakiyama, Koji Ide, Yoshinobu Tokyo Dent Coll Dept Anat Mihama Ku Chiba 2618502 Japan

This study clarifies the internal structure of the mandibular condyle in dentulous and edentulous jaws;three-dimensional observation and morphological measurements were performed using micro-CT. The materials used in this study were 14 mandibular condyles each removed from the dentulous and edentulous cadavers. Internal structures of the mandibular condyle were morphologically analyzed in three anatomical planes. Additionally, morphometrical observations of the trabecular bone structure at five regions of interest and cortical bone thickness were performed. The mandibular condyle of dentulous jaws consisted predominantly of regularly aligned plate-shaped trabeculae. Conversely, edentulous jaws were predominantly irregularly aligned narrow rod-shaped trabeculae. The results of morphometrical analysis showed that trabecular separation (***) in the central region of the mandibular condyle most closely reflected the difference between dentulous and edentulous jaws. These results suggest that, in edentulous jaws, reduced masticatory function due to tooth loss affects the internal structures of mandibular condyle.

关键词： Adult Aged Aged, 80 and over Bone Density/physiology Cadaver Dentition Female Humans Image Processing, Computer-Assisted/methods Imaging, Three-Dimensional/methods Jaw, Edentulous/pathology Jaw, Edentulous/radiography Male Mandible/pathology Mandible/radiography Mandibular Condyle/pathology Mandibular Condyle/radiography Middle Aged x-ray microtomography/methods

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