Human osteosarcoma cells in response to ELF-MF: Morphological remodeling compared to cell proliferation.

Purpose: The present study aimed to assess the effects of extremely low-frequency electromagnetic fields (ELF-MF) on structural changes of human osteosarcoma cells by analyzing the stained cytoskeleton for assessing the relationship between the fractal dimension parameter and proliferation rate of radiation-induced cells. Materials and Methods: In this study, 2-mT magnetic fields with various waveforms, including sinusoidal, triangular, and pulsed shapes, were employed to determine the biological effects of ELF-EMF on the human osteosarcoma MG-63 cell line. All experiments were performed in two modes: continuous exposure at 3 h and fractionated irradiations at 3 consecutive days. Afterward, the proliferation assay was implemented for assessing the cell proliferation in each group. Moreover, immunofluorescence staining and confocal imaging were performed to determine the cell shape index. Furthermore, fractal dimension analysis was carried out by processing morphological images. Results: The proliferation and shape index parameters of radiation-induced osteosarcomas significantly decreased compared with non-irradiated cells. In addition, fractal dimensions significantly increased following fractionated exposure at 3 consecutive days. Conclusions: Assessing the fractal dimensions can be considered as a new morphological index for the prognosis of the structural remodeling of human osteosarcoma cells in response to fractionated irradiation of ELF-MF. In addition, various waveforms induce a similar effect on morphological remodeling and cell proliferation.

Photobiomodulation Therapy Affects the Elastic Modulus, Cytoskeletal Rearrangement and Migration Capability of Human Osteosarcoma Cells.

Photobiomodulation (PBM) therapy utilizes low-power lasers to modulate the viability of living human cells and leads to changes in proliferation, differentiation, adhesion and gene expression, even though the rearrangement of cytoskeleton was not previously studied. The present study aims to evaluate the photobiological effects on the elastic behavior of human osteosarcoma cells (MG-63) and their morphological changes. Fluorescence staining, confocal imaging and atomic force microscopy (AFM) topography were performed to study the effects of PBM therapy with the exposure of 532 nm-25mW, 650 nm-3mW, 650 nm-150mW and 780 nm-70mW beams following the 5-min continuous irradiation. The area of each beam was 3.14cm2 with a source-surface distance of 20 cm. Besides the cell proliferation assessment, the migratory potential of MG-63 was determined with the wound healing technique. The results indicated an increase in stiffness and shape index of radiation-induced cells 24 h after exposure along with the obvious F-actins changes. But, cell stiffening was not observed 72 h after 532 nm laser irradiation. Also, a decrease in the migration rate was seen in all of the groups after 72 h of irradiation except cells treated with 532 nm wavelength. However, 532 nm laser beams increase the migratory potential 24 h after exposure. Within 72 h after irradiation, the cell proliferation was only affected by applying 532 nm and 650 nm-150mW laser beams. It was concluded that applying photobiomodulation with wavelengths of 650 nm (at both utilized powers) and 780 nm alters the migration capability and provides a quantitative description of cytoskeletal changes. Moreover, membrane stiffening can be considered as the biological marker of PBM treatments.

Assessing the short-term effects of radiotherapy on the shear modulus of the common carotid artery as a new biomarker of radiation-induced atherosclerosis.

PURPOSE: The purpose of this study was to investigate the incidence of short-term atherosclerosis in the common carotid arteries following radiotherapy. METHODS: The mean radiation dose to the arteries was 49.30±15.83 Gy. A computational ultrasound method was introduced to investigate the elastic modulus. Ultrasonography was performed 2-3 cm inferior to the bifurcation region before and after radiotherapy, and sequential images were extracted from a video of each artery. Instantaneous movement of the arterial wall in the radial and longitudinal directions was extracted by implementing the maximum gradient and block matching algorithms, respectively. RESULTS: There was a significant change in systolic blood pressure after radiotherapy (P=0.008). Irradiated arteries had significantly smaller systolic and end-diastolic diameters than non-irradiated arteries (P<0.001). The shear modulus was significantly different between irradiated and non-irradiated arteries (3.10±2.03 kPa vs. 1.38±0.98 kPa, P<0.001). The shear and Young moduli of radiation-induced arteries were 2.25±1.50 and 1.57±0.59 times higher than those of the pre-irradiation arteries. CONCLUSION: The arterial shear modulus can be considered as a new biomarker of radiationinduced atherosclerosis in the common carotid artery.

Attenuation correction in single-photon emission computed tomography for NURBS-based cardiac-torso phantom using dual-energy acquisition.

Single photon emission tomography is widely used to detect photons emitted from the patient. Some of these emitted photons suffer from scattering and absorption because of the attenuation occurred through their path in patient's body. Therefore, the attenuation is the most important problem in single-photon emission computed tomography (SPECT) imaging. Some of the radioisotopes emit gamma rays in different energy levels, and consequently, they have different counts and attenuation coefficients. Calculation of the parameters used in the attenuation equation N out=αNin = e- µ l Nin by mathematical methods is useful for the attenuation correction. Nurbs-based cardiac-torso (NCAT) phantom with an adequate attenuation coefficient and activity distribution is used in this study. Simulations were done using SimSET in 20-70 and 20-167 keV. A total of 128 projections were acquired over 360°. The corrected and reference images were compared using a universal image quality index (UIQI). The simulation repeated using NCAT phantom by SimSET. In the first group, no attenuation correction was used, but the Zubal coefficients were used for attenuation correction in the second image group. After the image reconstruction, a comparison between image groups was done using optimized UIQI to determine the quality of used reconstruction methods. Similarities of images were investigated by considering the average sinogram for every block size. The results showed that the proposed method improved the image quality. This study showed that simulation studies are useful tools in the investigation of nuclear medicine researches. We produced a nonattenuated model using Monte Carlo simulation method and compared it with an attenuated model. The proposed reconstruction method improved image resolution and contrast. Regional and general similarities of images could be determined, respectively, from acquired UIQI of small and large block sizes. Resulted curves from both small and large block sizes showed a good similarity between reconstructed and ideal images.

Dose-dependent 60Co γ-radiation Effects on Human Endothelial Cell Mechanical Properties.

Exposure to ionizing radiation is unavoidable for noncancerous cells during the external radiotherapy process. Increasing the dose delivery fraction times leads to increasing the endothelial cell damage. Vascular abnormalities are commonly associated with the alternation of endothelium biomechanical properties. The goal of the present study was to quantify the elastic and viscoelastic properties of human umbilical vein endothelial cells (HUVECs) using the micropipette aspiration technique in conjunction with a theoretical model while an 8 Gy dose was given in four fractions. Confocal imaging was performed for evaluation of cytoskeletal changes during fractionation 60Co radiotherapy. The results indicated an increase in elastic modulus from 29.87 ± 1.04 Pa to 46.69 ± 1.17 Pa while the fractional doses increased from 0 Gy to 8 Gy along with the obvious cytoskeletal changes. Moreover, in the creep behavior of radiated groups, a significant decrease was shown in the time constant and viscoelastic properties. On the other hand, it was observed that the change in the biomechanical properties of the cells while applying a single fraction of 8 Gy was not exactly the same as that in the properties of the radiation-exposed cells while delivering an 8 Gy dose at 2 Gy per fraction. The observed differences in the biomechanical behavior of endothelium provide a quantitative description of radiobiological effects for evaluating the dose-response relationship as a biological dosimetry procedure.

Radiation therapy affects the mechanical behavior of human umbilical vein endothelial cells.

Radiation therapy has been widely utilized as an effective method to eliminate malignant tumors and cancerous cells. However, subjection of healthy tissues and the related networks of blood vessels adjacent to the tumor area to irradiation is inevitable. The aim of this study was to investigate the consequent effects of fractionation radiotherapy on the mechanical characteristics of human umbilical vein endothelial cells (HUVECs) through alterations in cytoskeleton organization and cell and nucleus morphology. In order to simulate the clinical condition of radiotherapy, the HUVECs were exposed to the specific dose of 2 Gy for 1-4 times among four groups with incremental total dose from 2 Gy up to 8 Gy. Fluorescence staining was performed to label F-actin filaments and nuclei. Micropipette aspiration and standard linear solid model were employed to evaluate the elastic and viscoelastic characteristics of the HUVECs. Radiotherapy significantly increased cell elastic moduli. Due to irradiation, instantaneous and equilibrium Young's modulus were also increased. Radiotherapy diminished HUVECs viscoelastic behavior and shifted their creep compliance curves downward. Furthermore, gamma irradiation elevated the nuclei sizes and to a lesser extent the cells sizes resulting in the accumulation of F-actin filaments within the rest of cell body. Endothelial stiffening correlates with endothelial dysfunction, hence the results may be helpful when the consequent effects of radiotherapy are the focus of concern.

Hemodynamic analysis of radiation-induced damage in common carotid arteries by using color Doppler ultrasonography.

PURPOSE: The aim of this study was to assess vascular changes and blood flow abnormalities in the common carotid arteries of patients with head and neck cancers after external radiotherapy, using color Doppler ultrasonography. METHODS: We studied 24 patients treated with external radiotherapy for various head and neck cancers. In order to study the acute effects of irradiation on common carotid blood flow and arterial diameter changes, color Doppler ultrasonography parameters such as peak systolic velocity, end diastolic velocity, mean velocity, systolic-to-diastolic velocity (S/D) ratio, pulsatility index (PI), resistive index (RI), and instantaneous diameter changes were evaluated before and after external radiotherapy. Additionally, the blood volume flow (VF) values in the peak systolic and end diastolic phases, as well as mean velocity, were evaluated throughout three cardiac cycles using B-mode ultrasonic image processing. RESULTS: The findings showed significant changes in the S/D ratio, PI, and RI of the common carotid arteries before and after external radiotherapy (P<0.05). Moreover, a significant decrease in artery diameter and blood VF was observed after radiotherapy relative to the pretreatment values. A significant correlation was found between the blood VF values estimated using ultrasonic measurements and mathematical methods throughout three cardiac cycles. CONCLUSION: The hemodynamic parameters of the common carotid arteries changed during radiotherapy. These arterial changes may lead to late adverse effects of radiotherapy, such as ischemic strokes and ischemic attacks.

Evaluation of off-axis wedge correction factor using diode dosimeters for estimation of delivered dose in external radiotherapy.

An in vivo dosimetry system, using p-type diode dosimeters, was characterized for clinical applications of treatment machines ranging in megavoltage energies. This paper investigates two different models of diodes for externally wedged beams and explains a new algorithm for the calculation of the target dose at various tissue depths in external radiotherapy. The values of off-axis wedge correction factors were determined at two different positions in the wedged (toward the thick and thin edges) and in the non-wedged directions on entrance and exit surfaces of a polystyrene phantom in (60)Co and 6 MV photon beams. Depth transmission was defined on the entrance and exit surfaces to obtain the off-axis wedge correction factor at any depth. As the sensitivity of the diodes depends on physical characteristics [field size, source-skin distance (SSD), thickness, backscatter], correction factors were applied to the diode reading when measuring conditions different from calibration situations. The results indicate that needful correction factors for (60)Co wedged photons are usually larger than those for 6 MV wedged photon beams. In vivo dosimetry performed with the proposed algorithms at externally wedged beams has negligible probable errors (less than 0.5&) and is a reliable method for patient dose control.