3D strain map of axially loaded mouse tibia: a numerical analysis validated by experimental measurements

A combined experimental/numerical study was performed to calculate the 3D octahedral shear strain map in a mouse tibia loaded axially. This study is motivated by the fact that the bone remodelling analysis, in this in vivo mouse model should be performed at the zone of highest mechanical stimulus to maximise the measured effects. Accordingly, it is proposed that quantification of bone remodelling should be performed at the tibial crest and at the distal diaphysis. The numerical model could also be used to furnish a more subtle analysis as a precise correlation between local strain and local biological response can be obtained with the experimentally validated numerical model.     

Computational simulation of internal bone remodelling around dental implants: a sensitivity analysis

This study aimed to predict the distribution of bone trabeculae, as a density change per unit time, around a dental implant based on applying a selected mathematical remodelling model. The apparent bone density change as a function of the mechanical stimulus was the base of the applied remodelling model that describes disuse and overload bone resorption. The simulation was tested in a finite element model of a screw-shaped dental implant in an idealised bone segment. The sensitivity of the simulation to different mechanical parameters was investigated; these included element edge length, boundary conditions, as well as direction and magnitude of the implant loads. The alteration in the mechanical parameters had a significant influence on density distribution and model stability, in particular at the cortical bone region. The remodelling model could succeed to achieve trabeculae-like structure around osseointegrated dental implants. The validation of this model to a real clinical case is required.     

3D collagen cultures under well‐defined dynamic strain: A novel strain device with a porous elastomeric support

The field of mechanobiology has grown tremendously in the past few decades, and it is now well accepted that dynamic stresses and strains can impact cell and tissue organization, cell–cell and cell–matrix communication, matrix remodeling, cell proliferation and apoptosis, cell migration, and many other cell behaviors in both physiological and pathophysiological situations. Natural reconstituted matrices like collagen and fibrin are often used for three‐dimensional (3D) mechanobiology studies because they naturally form fibrous architectures and are rich in cell adhesion sites; however, they are physically weak and typically contain >99% water, making it difficult to apply dynamic stresses to them in a truly 3D context. Here we present a composite matrix and strain device that can support natural matrices within a macroporous elastic structure of polyurethane. We characterize this system both in terms of its mechanical behavior and its ability to support the growth and in vivo‐like behaviors of primary human lung fibroblasts cultured in collagen. The porous polyurethane was created with highly interconnected pores in the hundreds of µm size scale, so that while it did not affect cell behavior in the collagen gel within the pores, it could control the overall elastic behavior of the entire tissue culture system. In this way, a well‐defined dynamic strain could be imposed on the 3D collagen and cells within the collagen for several days (with elastic recoil driven by the polyurethane) without the typical matrix contraction by fibroblasts when cultured in 3D collagen gels. We show lung fibroblast‐to‐myofibroblast differentiation under 30%, 0.1 Hz dynamic strain to validate the model and demonstrate its usefulness for a wide range of tissue engineering applications. Biotechnol. Bioeng. 2009;103: 217–225. © 2008 Wiley Periodicals, Inc.     

Primary cilium mechanotransduction of tensile strain in 3D culture: Finite element analyses of strain amplification caused by tensile strain applied to a primary cilium embedded in a collagen matrix

Human adipose-derived stem cells (hASC) exhibit multilineage differentiation potential with lineage specification that is dictated by both the chemical and mechanical stimuli to which they are exposed. We have previously shown that 10% cyclic tensile strain increases hASC osteogenesis and cell-mediated calcium accretion. We have also recently shown that primary cilia are present on hASC and that chemically-induced lineage specification of hASC concurrently results in length and conformation changes of the primary cilia. Further, we have observed cilia length changes in hASC cultured within a collagen I gel in response to 10% cyclic tensile strain. We therefore hypothesize that primary cilia may play a key mechanotransduction role for hASC exposed to tensile strain. The goal of this study was to use finite element analysis (FEA) to determine strains occurring within the ciliary membrane in response to 10% tensile strain applied parallel, or perpendicular, to cilia orientation. To elucidate the mechanical environment experienced by the cilium, several lengths were modeled and evaluated based on cilia lengths measured on hASC grown under varied culture conditions. Principal tensile strains in both hASC and ciliary membranes were calculated using FEA, and the magnitude and location of maximum principal tensile strain determined. We found that maximum principal tensile strain was concentrated at the base of the cilium. In the linear elastic model, applying strain perpendicular to the cilium resulted in maximum strains within the ciliary membrane from 150% to 200%, while applying strain parallel to the cilium resulted in much higher strains, approximately 400%. In the hyperelastic model, applying strain perpendicular to the cilium resulted in maximum strains within the ciliary membrane around 30%, while applying strain parallel to the cilium resulted in much higher strains ranging from 50% to 70%. Interestingly, FEA results indicated that primary cilium length was not directly related to ciliary membrane strain. Rather, it appears that cilium orientation may be more important than cilium length in determining sensitivity of hASC to tensile strain. This is the first study to model the effects of tensile strain on the primary cilium and provides newfound insight into the potential role of the primary cilium as a mechanosensor, particularly in tensile strain and potentially a multitude of other mechanical stimuli beyond fluid shear.     

Left ventricular diastolic dysfunction in type 2 diabetes patients: a novel 2D strain analysis based on cardiac magnetic resonance imaging

Objective: This study was to develop a strain analysis method to evaluate the left ventricular (LV) functions in type 2 diabetic patients with an asymptomatic LV diastolic dysfunction. Methods: Two groups (10 asymptomatic type 2 diabetic subjects and 10 control ones) were considered. All of the subjects had normal ejection fraction values but impaired diastolic functions assessed by the transmitral blood flow velocity. For each subject, based on cardiac MRI, global indexes including LV volume, LV myocardial mass, cardiac index (CI), and transmitral peak velocity, were measured, and regional indexes (i.e., LV deformation, strain and strain rate) were calculated through an image-registration technology. Results: Most of the global indexes did not differentiate between the two groups, except for the CI, LV myocardial mass and transmitral peak velocity. While for the regional indexes, the global LV diastolic dysfunction of the diabetic indicated an increased strain (0.08?±?0.044 vs. ?0.031?±?0.077, p?=?0.001) and a reduced strain rate (1.834?±?0.909 vs. 3.791?±?2.394, p?=?0.033) compared to the controls, moreover, the local LV diastolic dysfunction reflected by the strain and strain rate varied, and the degree of dysfunction gradually decreased from the basal level to the apical level. Conclusions: The results showed that the strain and strain rates are effective to capture the subtle alterations of the LV functions, and the proposed method can be used to estimate the LV myocardial function based on cardiac MRI.     

Predicting the external formation of a bone fracture callus: an optimisation approach

The formation of a fracture callus in vivo tends to form in a structurally efficient manner distributing tissues where mechanical stimulus persists. Therefore, it is proposed that the formation of a fracture callus can be modelled in silico by way of an optimisation algorithm. This was tested by generating a finite element model of a transversal bone fracture embedded in a large tissue domain which was subjected to axial, bending and torsional loads. It was found that the relative fragment motion induced a compressive strain field in the early callus tissue which could be utilised to simulate the formation of external callus structures through an iterative optimisation process of tissue maintenance and removal. The phenomenological results showed a high level of congruence with in vivo healing patterns found in the literature. Consequently, the proposed strategy shows potential as a means of predicting spatial bone healing phenomena for pre-clinical testing.     

JULIDE: a software tool for 3D reconstruction and statistical analysis of autoradiographic mouse brain sections

In this article we introduce JULIDE, a software toolkit developed to perform the 3D reconstruction, intensity normalization, volume standardization by 3D image registration and voxel-wise statistical analysis of autoradiographs of mouse brain sections. This software tool has been developed in the open-source ITK software framework and is freely available under a GPL license. The article presents the complete image processing chain from raw data acquisition to 3D statistical group analysis. Results of the group comparison in the context of a study on spatial learning are shown as an illustration of the data that can be obtained with this tool.     

A comparison of stereology, structural rigidity and a novel 3D failure surface analysis method in the assessment of torsional strength and stiffness in a mouse tibia fracture model

In attempting to develop non-invasive image based measures for the determination of the biomechanical integrity of healing fractures, traditional μCT based measurements have been limited. This study presents the development and evaluation of a tool for assessment of fracture callus mechanical properties through determination of the geometric characteristics of the fracture callus, specifically along the surface of failure identified during destructive mechanical testing. Fractures were created in tibias of ten male mice and subjected to μCT imaging and biomechanical torsion testing. Failure surface analysis, along with previously described image based measures was calculated using the μCT image data, and correlated with mechanical strength and stiffness. Three-dimensional measures along the surface of failure, specifically the surface area and torsional rigidity of bone, were shown to be significantly correlating with mechanical strength and stiffness. It was also shown that surface area of bone along the failure surface exhibits stronger correlations with both strength and stiffness than measures of average and minimum torsional rigidity of the entire callus. Failure surfaces observed in this study were generally oriented at 45° to the long axis of the bone, and were not contained exclusively within the callus. This work represents a proof of concept study, and shows the potential utility of failure surface analysis in the assessment of fracture callus stability.     

Genetic variations and physical activity as determinants of limb bone morphology: an experimental approach using a mouse model

To gain insight into past human physical activity, anthropologists often infer functional loading history from the morphology of limb bone remains. It is assumed that, during life, loading had a positive, dose-dependent effect on bone structure that can be identified despite other effects. Here, we investigate the effects of genetic background and functional loading on limb bones using mice from an artificial selection experiment for high levels of voluntary wheel running. Growing males from four replicate high runner (HR) lines and four replicate nonselected control (C) lines were either allowed or denied wheel access for 2 months. Using μCT, femoral morphology was assessed at two cortical sites (mid-diaphysis, distal metaphysis) and one trabecular site (distal metaphysis). We found that genetic differences between the linetypes (HR vs. C), between the replicate lines within linetype, and between individuals with and without the so-called "mini-muscle" phenotype (caused by a Mendelian recessive gene that halves limb muscle mass) gave rise to significant variation in nearly all morphological indices examined. Wheel access also influenced femoral morphology, although the functional response did not generally result in enhanced structure. Exercise caused moderate periosteal enlargement, but relatively greater endocortical expansion, resulting in significantly thinner cortices and reduced bone area in the metaphysis. The magnitude of the response was independent of distance run. Mid-diaphyseal bone area and area moments, and trabecular morphology, were unaffected by exercise. These results underscore the strong influence of genetics on bone structure and the complexity by which mechanical stimuli may cause alterations in it.     

Mechanical response comparison in an implant overdenture retained by ball attachments on conventional regular and mini dental implants: a finite element analysis

This study investigates the bone/implant mechanical responses in an implant overdenture retained by ball attachments on two conventional regular dental implants (RDI) and four mini dental implants (MDI) using finite element (FE) analysis. Two FE models of overdentures retained by RDIs and MDIs for a mandibular edentulous patient with validation within 6% variation errors were constructed by integrating CT images and CAD system. Bone grafting resulted in 2 mm thickness at the buccal side constructed for the RDIs-supported model to mimic the bone augmentation condition for the atrophic alveolar ridge. Nonlinear hyperelastic material and frictional contact element were used to simulate characteristic of the ball attachment-retained overdentures. The results showed that a denture supported by MDIs presented higher surrounding bone strains than those supported by RDIs under different load conditions. Maximum bone micro strains were up to 6437/2987 and 13323/5856 for MDIs/RDIs under single centric and lateral contacts, respectively. Corresponding values were 4429/2579 and 9557/5774 under multi- centric and lateral contacts, respectively. Bone micro strains increased 2.06 and 1.96-folds under single contact, 2.16 and 2.24-folds under multiple contacts for MDIs and RDIs when lateral to axial loads were compared. The maximum RDIs and MDIs implant stresses in all simulated cases were found by far lower than their yield strength. Overdentures retained using ball attachments on MDIs in poor edentulous bone structure increase the surrounding bone strain over the critical value, thereby damaging the bone when compared to the RDIs. Eliminating the occlusal single contact and oblique load of an implant-retained overdenture reduces the risk for failure.     

Fate and behaviour of a seed-applied Pseudomonas brassicacearum strain in a winter wheat field trial,as determined by analysis with SCAR markers

The fate and behaviour of the seed-applied biocontrol strain Pseudomonas brassicacearum MA250 in a field trial with winter wheat was determined using sequence-characterised amplified region (SCAR) markers. Samples of belowground plant parts from healthy and withered (due to snow mould infection) seedlings were collected approximately one and seven months after sowing, which was performed in early autumn. DNA was extracted from roots and remaining parts of seeds with adhering soil, and the abundance of the strain was determined in quantitative real-time PCR (qPCR) assays. The results show that the introduced strain persisted over the whole trial-period of seven months. On termination of the trial (after seven months) the belowground plant parts of each plant housed 10⁶–10⁷ cells, substantially less than the original approximately 10⁹ cells inoculated onto the seed. In healthy seedlings, there was a shift in cell numbers from seeds to roots between the samplings, suggesting colonisation of the roots during this time. The results show that with sufficient attention given to analytical control measures and the possibility of resident background populations, SCAR markers in combination with qPCR provide valuable information regarding the fate and behaviour of biocontrol micro-organisms under field conditions.     

3-苯氧基苯甲酸降解菌Sphingomonas sp. SC-1降解苯酚环境条件及其降解中间产物的研究

【目的】探究高效降解3-苯氧基苯甲酸(3-Phenoxybenzoic acid,3-PBA)的鞘氨醇单胞菌(Sphingomonas sp.) SC-1对苯酚的降解特性。【方法】采用HPLC测定微生物降解体系中苯酚残留量,考察环境条件对菌株SC-1降解苯酚的影响;分析不同培养时间苯酚降解体系混合样品的HPLC谱图,确定其降解中间产物。【结果】菌株SC-1能在基础盐培养基中以苯酚为唯一碳源和能源生长,在初始pH 7.0、30 °C条件下,24 h可完全降解100 mg/L苯酚;Cu2+、Ba2+、Mn2+等对其降解苯酚有不同程度的抑制作用;HPLC谱图分析,初步确定邻苯二酚是菌株SC-1降解苯酚的中间产物,且该菌株可在48 h内完全降解100 mg/L邻苯二酚。【结论】菌株SC-1对苯酚及邻苯二酚均有较强的降解能力,为完善3-PBA的降解途径及污染3-PBA或含酚废水或含酚农药残留的降解提供了数据参考。     

Numerical simulation and PEPT measurements of a 3D conical helical-blade mixer: a high potential solids mixer for solid-state fermentation

Helical-blade solids mixers have a large potential as bioreactors for solid-state fermentation (SSF). Fundamental knowledge of the flow and mixing behavior is required for robust operation of these mixers. In this study predictions of a discrete particle model were compared to experiments with colored wheat grain particles and positron emission particle tracking (PEPT) measurements. In the discrete particle model individual movements of particles were calculated from interaction forces. It was concluded that the predicted overall flow behavior matched well with the PEPT measurements. Differences between the model predictions and the experiments with wheat grains were found to be due to the assumption that substrate particles were spherical, which was in the model. Model simulations and experiments with spherical green peas confirmed this. The mixing in the helical-blade mixer could be attributed to (1) the transport of particles up and down in the interior of the mixer, and (2) dispersion or micro-mixing of particles in the top region of the mixer. It appeared that the mixing rate scaled linearly with the rotation rate of the blade, although the average particle velocity did not scale proportionally. It may be that the flow behavior changes as a function of the rotation rate (e.g., changing thickness of the top region); further study is required to confirm this. To increase the mixing performance of the mixer, a larger blade or a change in the shape of the mixer (larger top surface/volume ratio) is recommended.     

Immediate induction of a 45 K secreted glycoprotein by serum and growth factors in quiescent mouse 3T3 cells : Two-dimensional gel analysis

Stimulation of quiescent 3T3 cells by serum dramatically induces the synthesis of a group of secreted polypeptides with a molecular weight (MW) of 45 K (p45 A, B, C, D). The synthesis of these polypeptides increases 10-fold during the first 2 h. Cycloheximide superinduces the 45 K polypeptides and actinomycin D (actD) blocks completely their induction by serum. Peptide mapping analysis and pulse-chase experiments revealed that p45 A is a precursor of polypeptides p45 B, C, D. Tunicamycin treatment inhibits the synthesis of all four polypeptides but a new related protein appears, p-p45, the unglycosylated precursor. In the presence of tunicamycin, p-p45 is also found in the medium, demonstrating that glycosylation is not essential for the secretion. In vitro translation experiments show that the levels of p45 mRNA present in stimulated cells are severalfold higher than that of non-stimulated cells. Purified growth factors, such as platelet-derived growth factor (PDGF), fibroblast growth factor (FGF) and epidermal growth factor (EGF) induce the synthesis of p45 in quiescent cells.     

Cartesian stiffness for wrist joints: analysis on the Lie group of 3D rotations and geometric approximation for experimental evaluation

This paper is concerned with the analysis and the numerical evaluation from experimental measurements of the static, Cartesian stiffness of wrist joints, in particular the human wrist. The primary aim is to extend from Euclidean spaces to so(3), the group of rigid body rotations, previous methods for assessing the end-point stiffness of the human arm, typically performed via a robotic manipulandum. As a first step, the geometric definition of Cartesian stiffness from current literature is specialised to the group so(3). Emphasis is placed on the choice of the unique, natural, affine connection on so(3) which guarantees symmetry of the stiffness matrix in presence of conservative fields for any configuration, also out of equilibrium. As the main contribution of this study, a coordinate-independent approximation based on the geometric notion of geodesics is proposed which provides a working equation for evaluating stiffness directly from experimental measurements. Finally, a graphical representation of the stiffness is discussed which extends the ellipse method often used for end-point stiffness visualisation and which is suitable to compare stiffness matrices evaluated at different configurations.     

Inhibition of proliferation and induction of differentiation of osteoblastic cells by a novel 1,25-dihydroxyvitamin D3 analog with an extensively modified side chain (CB1093)

1,25-Dihydroxyvitamin D₃ (1,25D) is involved in the regulation of proliferation and differentiation of a variety of cell types including cancer cells. In recent years, numerous new vitamin D₃ analogs have been developed in order to obtain favorable therapeutic properties. The effects of a new 20-epi analog, CB1093 (20-epi-22-ethoxy-23-yne-24a,26a,27a-trihomo-1α,25(OH)₂D₃), on the proliferation and differentiation of human MG-63 osteosarcoma cell line were compared here with those of the parent compound 1,25D. Proliferation of the MG-63 cells was inhibited similarly by 22%, 50% and 59% after treatment with 0.1 μM 1,25D or CB1093 for 48 h, 96 h, and 144 h, respectively. In transfection experiments, the compounds were equipotent in stimulating reporter gene activity under the control of human osteocalcin gene promoter. In cell culture experiments, however, CB1093 was more potent than 1,25D at low concentrations and more effective for a longer period of time in activating the osteocalcin gene expression at mRNA and protein levels. Also, a 6-h pretreatment and subsequent culture for up to 120 h without 1,25D or CB1093 yielded higher osteocalcin mRNA and protein levels with analog-treated cells than with 1,25D-treated cells. The electrophoretic mobility shift assay (EMSA) revealed stronger VDR-VDRE binding with analog-treated MG-63 cells than with 1,25D-treated cells. The differences in the DNA binding of 1,25D-bound vs. analog-bound VDR, however, largely disappeared when the binding reactions were performed with recombinant hVDR and hRXRβ proteins. These results demonstrate that the new analog CB1093 was equally or even more effective than 1,25D in regulating all human osteosarcoma cell functions ranging from growth inhibition to marker gene expression and that the differences in effectivity most probably resulted from interactions of the hVDR:hRXRβ-complex with additional nuclear proteins. J. Cell. Biochem. 70:414–424, 1998. © 1998 Wiley-Liss, Inc.