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Great controversy exists regarding the biologic responses of osteoblasts to X-ray irradiation, and the mechanisms are poorly understood. In this study, the biological effects of low-dose radiation on stimulating osteoblast proliferation, differentiation and fracture healing were identified using in vitro cell culture and in vivo animal studies. First, low-dose (0.5 Gy) X-ray irradiation induced the cell viability and proliferation of MC3T3-E1 cells. However, high-dose (5 Gy) X-ray irradiation inhibited the viability and proliferation of osteoblasts. In addition, dynamic variations in osteoblast differentiation markers, including type I collagen, alkaline phosphatase, Runx2, Osterix and osteocalcin, were observed after both low-dose and high-dose irradiation by Western blot analysis. Second, fracture healing was evaluated via histology and gene expression after single-dose X-ray irradiation, and low-dose X-ray irradiation accelerates fracture healing of closed femoral fractures in rats. In low-dose X-ray irradiated fractures, an increase in proliferating cell nuclear antigen (PCNA)-positive cells, cartilage formation and fracture calluses was observed. In addition, we observed more rapid completion of endochondral and intramembranous ossification, which was accompanied by altered expression of genes involved in bone remodeling and fracture callus mineralization. Although the expression level of several osteoblast differentiation genes was increased in the fracture calluses of high-dose irradiated rats, the callus formation and fracture union were delayed compared with the control and low-dose irradiated fractures. These results reveal beneficial effects of low-dose irradiation, including the stimulation of osteoblast proliferation, differentiation and fracture healing, and highlight its potential translational application in novel therapies against bone-related diseases.

Microdamage occurs in bone through repeated and excessive loading. Accumulation of microdamage weakens bone, leading to a loss of strength, stiffness and energy dissipation in the tissue. Imaging techniques used to examine microdamage have typically been limited to the microscale. In the current study microdamage was examined at the nanoscale using transmission x-ray microscopy with an x-ray negative stain, lead-uranyl acetate. Microdamage was generated in notched and unnotched beams of sheep cortical bone (2×2×20 mm), with monotonic and fatigue loading. Bulk sections were removed from beams and stained with lead-uranyl acetate to identify microdamage. Samples were sectioned to 50 microns and imaged using transmission x-ray microscopy producing projection images of microdamage with nanoscale resolution. Staining indicated microdamage occurred in both the tensile and compressive regions. A comparison between monotonic and fatigue loading indicated a statistically significant greater amount of stain present in fatigue loaded sections. Microdamage occurred in three forms: staining to existing bone structures, cross hatch damage and a single crack extending from the notch tip. Comparison to microcomputed tomography demonstrated differences in damage morphology and total damage between the microscale and nanoscale. This method has future applications for understanding the underlying mechanisms for microdamage formation as well as three-dimensional nanoscale examination of microdamage.

Made available in DSpace on 2018-11-26T16:32:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-04-21 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Digital radiographic imaging is increasing in veterinary practice. The use of radiation demands responsibility to maintain high image quality. Low doses are necessary because workers are requested to restrain the animal. Optimizing digital systems is necessary to avoid unnecessary exposure, causing the phenomenon known as dose creep. Homogeneous phantoms are widely used to optimize image quality and dose. We developed an automatic computational methodology to classify and quantify tissues (i.e., lung tissue, adipose tissue, muscle tissue, and bone) in canine chest computed tomography exams. The thickness of each tissue was converted to simulator materials (i.e., Lucite, aluminum, and air). Dogs were separated into groups of 20 animals each according to weight. Mean weights were 6.5 +/- 2.0 kg, 15.0 +/- 5.0 kg, 32.0 +/- 5.5 kg, and 50.0 +/- 12.0 kg, for the small, medium, large, and giant groups, respectively. The one-way analysis of variance revealed significant differences in all simulator material thicknesses (p < 0.05) quantified between groups. As a result, four phantoms were constructed for dorsoventral and lateral views. In conclusion, the present methodology allows the development of phantoms of the canine chest and possibly other body regions and/or animals. The proposed phantom is a practical tool that may be employed in future work to optimize veterinary X-ray procedures. Sao Paulo State Univ, Dept Phys & Biophys, Biosci Inst Botucatu, Sao Paulo, Brazil Sao Paulo State Univ, Dept Trop Dis & Diagnost Imaging, Botucatu Med Sch, Sao Paulo, Brazil Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Anim Reprod & Vet Radiol, Sao Paulo, Brazil Brasilian Ctr Phys Res, Rio De Janeiro, Brazil Sao Paulo State Univ, Dept Phys & Biophys, Biosci Inst Botucatu, Sao Paulo, Brazil Sao Paulo State Univ, Dept Trop Dis & Diagnost Imaging, Botucatu Med Sch, Sao Paulo, Brazil Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Anim Reprod & Vet Radiol, Sao Paulo, Brazil

Imaging of cellular structure and extended tissue in biological materials requires nanometer resolution and good sample penetration, which can be provided by current full-field transmission X-ray microscopic techniques in the soft and hard X-ray regions. The various capabilities of full-field transmission X-ray microscopy (TXM) include 3D tomography, Zernike phase contrast, quantification of absorption, and chemical identification via X-ray fluorescence and X-ray absorption near edge structure imaging. These techniques are discussed and compared in light of results from the imaging of biological materials including microorganisms, bone and mineralized tissue, and plants, with a focus on hard X-ray TXM at ≤ 40-nm resolution. © 2010 Wiley-Liss, Inc.