Novel model of restricted mobility induced osteopenia in zebrafish

Immobilization, such as prolonged bed rest, is a risk factor for bone loss in humans. Motivated by the emerging utility of zebrafish (Danio rerio) as an animal of choice for the study of musculoskeletal disease, here we report a model of restricted mobility induced osteopenia in adult zebrafish. Aquatic tanks with small cubical compartments to restrict the movement and locomotion of single fish were designed and fabricated for this study. Adult zebrafish were divided into two groups: a normal control (CONT) and a restricted mobility group (RMG) (18 fish/group). Six fish from each group were euthanized on days 14, 21 and 35 of the movement restriction. By using microcomputed tomography (micro-CT), we assessed bone volume/tissue volume (BV/TV) and bone density in the whole skeleton of the fish. Furthermore, we assessed skeletal shape in the vertebrae (radius, length, volume, neural and haemal arch aperture areas, neural and haemal arch angle, and thickness of the intervertebral space), single vertebra bone volume and bone density. Movement restriction significantly decreased vertebral skeletal parameters such as radius, length, volume, arch aperture areas and angles as well as the thickness of the intervertebral space in RMG. Furthermore, restricted mobility significantly (P < 0.001) decreased BV/TV and bone density as compared to the CONT group, starting as early as 14 days. By analysing zebrafish from CONT and RMG, we show that micro-CT imaging is a sensitive method to quantify distinct skeletal properties in zebrafish. We further defined the micro-CT parameters which can be used to examine the effects of restricted mobility on the skeleton of the fish. Our findings propose a rapid and effective osteopenia “stabulation” model, which could be used widely for osteoporosis drug screening.     

Intravoxel bone micromechanics for microCT-based finite element simulations

While micro-FE simulations have become a standard tool in computational biomechanics, the choice of appropriate material properties is still a relevant topic, typically involving empirical grey value-to-elastic modulus relations. We here derive the voxel-specific volume fractions of mineral, collagen, and water, from tissue-independent bilinear relations between mineral and collagen content in extracellular bone tissue (J. Theor. Biol. 287: 115, 2011), and from the measured X-ray attenuation information quantified in terms of grey values. The aforementioned volume fractions enter a micromechanics representation of bone tissue, as to deliver voxel-specific stiffness tensors. In order to check the relevance of this strategy, we convert a micro Computer Tomograph of a mouse femur into a regular Finite Element mesh, apply forces related to the dead load of a standing mouse, and then compare simulation results based on voxel-specific heterogeneous elastic properties to results based on homogeneous elastic properties related to the spatial average over the solid bone matrix compartment, of the X-ray attenuation coefficients. The element-specific strain energy density illustrates that use of homogeneous elastic properties implies overestimation of the organ stiffness. Moreover, the simulation reveals large tensile normal stresses throughout the femur neck, which may explain the mouse femur neck's trabecular morphology being quite different from the human case, where the femur neck bears compressive forces and bending moments.     

CAD–CAM prosthetically guided bone regeneration using preformed titanium mesh for the reconstruction of atrophic maxillary arches

The protocol presented here is intended to minimise the intervention in bone reconstruction surgery when severe atrophy or deformity is present in the maxillary arches. A patient underwent augmentation of an atrophic maxillary arch using titanium mesh and particulate autogenous plus bovine demineralised bone. After computed tomography data elaboration, computer-aided design and computer-aided machining were used to plan the augmentation of bone volume to improve the implant position needed to support the final dental prosthesis. The augmented maxilla was rapidly prototyped in plastic, and the titanium mesh was tested on this model before the surgical intervention. Then, the preformed titanium mesh was implanted in the maxillary arch with bone grafting. The bone was augmented relative to the position of the implants for the definitive fixed implant-supported rehabilitation. The protocol presented here is a viable, reproducible way to determine the correct bone augmentation for the final implant-supported prosthetic rehabilitation.     

Computed tomography-based joint locations affect calculation of joint moments during gait when compared to scaling approaches

Hip joint moments are an important parameter in the biomechanical evaluation of orthopaedic surgery. Joint moments are generally calculated using scaled generic musculoskeletal models. However, due to anatomical variability or pathology, such models may differ from the patient's anatomy, calling into question the accuracy of the resulting joint moments. This study aimed to quantify the potential joint moment errors caused by geometrical inaccuracies in scaled models, during gait, for eight test subjects. For comparison, a semi-automatic computed tomography (CT)-based workflow was introduced to create models with subject-specific joint locations and inertial parameters. 3D surface models of the femora and hemipelves were created by segmentation and the hip joint centres and knee axes were located in these models. The scaled models systematically located the hip joint centre (HJC) up to 33.6 mm too inferiorly. As a consequence, significant and substantial peak hip extension and abduction moment differences were recorded, with, respectively, up to 23.1% and 15.8% higher values in the image-based models. These findings reaffirm the importance of accurate HJC estimation, which may be achieved using CT- or radiography-based subject-specific modelling. However, obesity-related gait analysis marker placement errors may have influenced these results and more research is needed to overcome these artefacts.     

Image assessment of MSCT and CBCT scans for rapid maxillary expansion: a pilot study

Objectives: The accuracy of cone-beam technique, cone-beam computed tomography (CBCT), compared with that of the multislice spiral CT (MSCT), for image-based linear measurements of midpalatal suture was assessed.

Material and Methods: Two measurements were performed by one investigator on the dry skull by using one digital caliper and in the axial cuts by using software. A 2D object-based image registration process was applied to determine the best affine transformation that maps a 2D input image (CBCT) in a reference image (MSCT).

Results: The value of the intraclass correlation coefficient was approximately 0.9%. The results suggest that differences between two scanners did not exist (observation 1, p = 0.964 and observation 2, p = 0.795). With regard to the dry skull and the image, the significance probabilities equaled zero (observation 1, p = 0.002 and observation 2, p = 0.004), therefore, indicating significant differences.

Conclusion: Measurements acquired in the images were similar and these findings contribute to stimulate the use of CBCT for evaluation of the maxillary expansion procedure.     

Synthesis and in vitro cellular uptake of 11C-labeled 5-aminolevulinic acid derivative to estimate the induced cellular accumulation of protoporphyrin IX

Protoporphyrin IX (PpIX) accumulation induced by exogenous 5-aminolevulinic acid (ALA) in tumors affects the therapeutic efficacy of ALA-based photodynamic and sonodynamic therapies. To develop a new imaging probe to estimate the ALA-induced PpIX accumulation, ¹¹C-labeled ALA analog (4), an ALA-dehydratase inhibitor, was radiosynthesized via ¹¹C-methylation of a Schiff-base-activated precursor in the presence of tetrabutylammonium fluoride, followed by the hydrolysis of ester and imine groups. The cellular uptake of 4 linearly increased with time and was inhibited by ALA and other transporter competitors. Monitoring analog 4 with positron emission tomography might be useful to estimate the ALA-induced PpIX accumulation in tumors.     

Towards tropomyosin-related kinase B (TrkB) receptor ligands for brain imaging with PET: Radiosynthesis and evaluation of 2-(4-[18F]fluorophenyl)-7,8-dihydroxy-4H-chromen-4-one and 2-(4-([N-methyl-11C]-dimethylamino)phenyl)-7,8-dihydroxy-4H-chromen-4-one

The interaction of tropomyosin-related kinase B (TrkB) with the cognate ligand brain-derived neurotrophic factor (BDNF) mediates fundamental pathways in the development of the nervous system. TrkB signaling alterations are linked to numerous neurodegenerative diseases and conditions. Herein we report the synthesis, biological evaluation and radiosynthesis of the first TrkB radioligands based on the recently identified 7,8-dihydroxyflavone chemotype. 2-(4-[¹⁸F]fluorophenyl)-7,8-dihydroxy-4H-chromen-4-one ([¹⁸F]10b) was synthesized in high radiochemical yields via an efficient S_NAr radiofluorination involving a para-Michael acceptor substituted aryl followed by BBr₃-promoted double demethylation. Selective N-[¹¹C]methylation afforded 2-(4-([N-methyl-¹¹C]-dimethylamino)phenyl)-7,8-dihydroxy-4H-chromen-4-one ([¹¹C]10c) from the fully deprotected catechol-bearing normethyl precursor 13 with [¹¹C]MeOTf. In vitro autoradiography of [¹⁸F]10b with transverse rat brain sections revealed high specific binding in the cortex, striatum, hippocampus and thalamus in accordance with expected TrkB distribution. Blockade experiments with both 7,8-dihydroxyflavone (1a) and TrkB cognate ligand, BDNF, led to decreases of 80% and 85% of radioligand binding strongly supporting the hypothesis that 7,8-dihydroxyflavones exert their effect on TrkB phosphorylation via direct TrkB extracellular domain (ECD) binding. Positron emission tomography (PET) studies revealed that [¹⁸F]10b and [¹¹C]10c brain uptake is minimal and that they are rapidly eliminated from the plasma (effective plasma half-life 5–10 min) via hepatic secretion. Nevertheless, the high specific binding and TrkB specificity derived from in vitro experiments suggests that the 7,8-disubstituted flavone chemotype represents a promising scaffold for the development of TrkB radiotracers for PET.     

Carbon‐11 radiolabeling of an oligopeptide containing tryptophan hydrochloride via a Pictet‐Spengler reaction using carbon‐11 formaldehyde

A procedure for the synthesis of a¹¹C‐labeled oligopeptide containing [1‐¹¹C]1,2,3,4‐tetrahydro‐β‐carboline‐3‐carboxylic acid ([1‐¹¹C]Tpi) from the corresponding Trp?HCl‐containing peptides has been developed involving a Pictet‐Spengler reaction with [¹¹C]formaldehyde. The synthesis of [1‐¹¹C]Tpi from Trp and [¹¹C]formaldehyde was examined as a model reaction with the aim of developing a facile and effective method for the labeling of peptides with carbon‐11. The Pictet‐Spengler reaction of Trp and [¹¹C]formaldehyde in acidic media (TsOH or HCl) afforded the desired [1‐¹¹C]Tpi in a moderate radiochemical yield. Herein, the application of a Pictet‐Spengler reaction to an aqueous solution of Trp?HCl gave the desired product with a radiochemical yield of 45.2%. The RGD peptide cyclo[Arg‐Gly‐Asp‐D‐Tyr‐Lys] was then selected as a substrate for the labeling reaction with [¹¹C]formaldehyde. The radiolabeling of a Trp?HCl‐containing RGD peptide using the Pictet‐Spengler reaction was successful. Furthermore, the remote‐controlled synthesis of a [1‐¹¹C]Tpi‐containing RGD peptide was attempted by using an automatic production system to generate [¹¹C]CH₃I. The radiochemical yield of the [1‐¹¹C]Tpi‐containing RGD at the end of synthesis (EOS) was 5.9 ± 1.9% (n = 4), for a total synthesis time of about 35 min. The specific activity was 85.7 ± 9.4 GBq/µmol at the EOS. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.