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Micro- and macrorheological properties of actin networks effectively cross-linked by depletion forces.

The structure and rheology of cytoskeletal networks are regulated by actin binding proteins. Aside from these specific interactions, depletion forces can also alter the properties of cytoskeletal networks. Here we demonstrate that the addition of poly(ethylene glycol) (PEG) as a depletion agent results not only in severe structural changes, but also in alterations in mechanical properties of actin solutions. In the plateau of the elastic modulus two regimes can be distinguished by micro and macrorheological methods. In the first, the elastic modulus increases only slightly with increasing depletion agent, whereas above a critical concentration c*, a strong increase of cPEG6k3.5 is observed in a distinct second regime. Microrheological data and electron microscopy images show a homogenous network of actin filaments in the first regime, whereas at higher PEG concentrations a network of actin bundles is observed. The concentration dependence of the plateau modulus G0, the shift in entanglement time taue, and the nonlinear response indicate that below c* the network becomes effectively cross-linked, whereas above c* G0(cPEG6k) is primarily determined by the network of bundles that exhibits a linearly increasing bundle thickness.

Actin Cytoskeleton↗

[Method of measuring the modulus of elasticity of human muscle tissue].

An experimental method is described for estimating the mechanical properties of human soft tissues by calculating Young's modulus which was defined according to the slope of the hysteresis curve of the test subject, reflecting the dependence of its deformation on the force applied. The main causes of numerous difficulties in obtaining such data and the ways for their overcoming are considered. Experiments were made on the biceps in the isometric mode under different degrees of internal tension. It has been shown that the higher the physical development of the test subject the less the magnitude of Young's modulus. The dependence of its magnitude on the degree of muscular tension is sublinear in character in the group with good physical development and superlinear in that with poor physical development.

Adult↗

Connections between single-cell biomechanics and human disease states: gastrointestinal cancer and malaria.

We investigate connections between single-cell mechanical properties and subcellular structural reorganization from biochemical factors in the context of two distinctly different human diseases: gastrointestinal tumor and malaria. Although the cell lineages and the biochemical links to pathogenesis are vastly different in these two cases, we compare and contrast chemomechanical pathways whereby intracellular structural rearrangements lead to global changes in mechanical deformability of the cell. This single-cell biomechanical response, in turn, seems to mediate cell mobility and thereby facilitates disease progression in situations where the elastic modulus increases or decreases due to membrane or cytoskeleton reorganization. We first present new experiments on elastic response and energy dissipation under repeated tensile loading of epithelial pancreatic cancer cells in force- or displacement-control. Energy dissipation from repeated stretching significantly increases and the cell's elastic modulus decreases after treatment of Panc-1 pancreatic cancer cells with sphingosylphosphorylcholine (SPC), a bioactive lipid that influences cancer metastasis. When the cell is treated instead with lysophosphatidic acid, which facilitates actin stress fiber formation, neither energy dissipation nor modulus is noticeably affected. Integrating recent studies with our new observations, we ascribe these trends to possible SPC-induced reorganization primarily of keratin network to perinuclear region of cell; the intermediate filament fraction of the cytoskeleton thus appears to dominate deformability of the epithelial cell. Possible consequences of these results to cell mobility and cancer metastasis are postulated. We then turn attention to progressive changes in mechanical properties of the human red blood cell (RBC) infected with the malaria parasite Plasmodium falciparum. We present, for the first time, continuous force-displacement curves obtained from in-vitro deformation of RBC with optical tweezers for different intracellular developmental stages of parasite. The shear modulus of RBC is found to increase up to 10-fold during parasite development, which is a noticeably greater effect than that from prior estimates. By integrating our new experimental results with published literature on deformability of Plasmodium-harbouring RBC, we examine the biochemical conditions mediating increases or decreases in modulus, and their implications for disease progression. Some general perspectives on connections among structure, single-cell mechanical properties and biological responses associated with pathogenic processes are also provided in the context of the two diseases considered in this work.

Animals↗

Quantitative constitutive behaviour and viscoelastic properties of fresh flexor tendons.

The objective of this study was to obtain detailed high quality experimental data under well-controlled test conditions in order to quantify tendon viscoelastic behaviour and provide an experimental basis for large deformation mathematical modelling. Eighty-six fresh chicken flexor digitorum profundus (FDP) tendons were mechanically tested using an Instron 5565 universal testing machine and a Bioplus bath containing physiological saline solution. At low strain rates (<or=0.001s(-1)), no strain rate dependence was found and the value of the elastic modulus was 324+/-17 MPa. At medium strain rates (0.003s(-1) - 0.1s(-1)), the elastic stiffness increased with increasing strain rate. For example, the values of the elastic modulus were 427+/-10, 653+/-21 and 837+/-11 MPa for strain rates of 0.006s(-1), 0.012s(-1) and 0.05s(-1) respectively. A series of stress relaxation experiments at different levels of strain were conducted. The higher the initial stress level, the faster the stress relaxed. For all stress relaxation tests, the relationship of normalised relaxation function G(t) against log time (ln(t)) was approximately linear (R2 = 0.935 +/- 0.028) and this supports Fung's quasi-linear viscoelasticity (QLV) hypothesis. Some preliminary work on bovine flexor tendons using a video extensometer is also reported.

Animals↗

Opening and closing characteristics of the aortic valve after valve-sparing procedures using a new aortic root conduit.

BACKGROUND: The durability of aortic valve-sparing procedures is negatively affected by increased leaflet stress in the absence of normally shaped sinuses of Valsalva. We compared valve motion after remodeling procedures using a standard conduit and a specifically designed aortic root conduit. METHODS: Echocardiographic studies of the aortic valve dynamics were performed in 14 patients after remodeling of the aortic root (7 standard conduits, group A; 7 new conduits, group B) and in 7 controls (group C). Opening and closing leaflet velocities and percent of slow closing leaflet displacement were measured. Root distensibility and the pressure strain of the elastic modulus were measured at all root levels. RESULTS: Root distensibility and the pressure strain of the elastic modulus were different in group A and B only at the sinuses (p < 0.001). Opening and closing leaflet velocities were not different among groups. Slow closing leaflet displacement was markedly more evident in group B patients (24.2%+/-1.9% versus 2.5%+/-1.9% in group A, p < 0.001) and similar to controls (22.1%+/-7.9%). CONCLUSIONS: The new conduit guarantees dynamic features of the aortic valve leaflets superior to those obtained with standard conduits and more similar to normal subjects.

Aged↗

Role of venous elasticity in the development of varicose veins.

A reduction in the 'elasticity' of the venous system has been proposed as a precursor of venous insufficiency, but the concept remains controversial. This study was designed to develop a method of assessing venous elasticity, and to use this method to investigate the aetiology of varicose veins. Simultaneous measurements of calf volume (determined using strain gauge plethysmography) and venous pressure (obtained via a dorsal foot vein) were made during venous occlusion plethysmography. The elastic modulus, K, defined as stress/strain when the veins are full, was calculated from the pressure/volume relationship. The elastic modulus was determined in 19 normal legs, 33 legs with superficial venous insufficiency, 16 legs with deep venous insufficiency, and 18 legs of a high risk group of volunteers or patients without varicose veins but with a strong history of factors associated with their development. The results showed a clear difference in elasticity between normal limbs and limbs with varicose veins, and also between normal limbs and high risk limbs. These results support the hypothesis that reduced elasticity has a role in the development of varicose veins and precedes the onset of valvular incompetence.

Elasticity↗

Cellular and membrane deformability of red blood cells in preterm infants with and without growth retardation.

Membrane and whole red cell deformability are essential for effective blood flow, oxygen uptake and release. Whole cell deformability (rheoscope with shear stresses of 0.6-30 Pa) and membrane shear elastic modulus (flow channel) of red blood cells (RBC) were studied in appropriate and small for gestational age preterm infants (28-32 wk of gestation), in healthy full-term neonates and in adults. No significant differences were found between the two groups of preterm infants. The preterm infants showed significantly increased RBC deformation in the rheoscope when compared to the full-term neonates who, in turn, showed more RBC deformation than adults. Correspondingly, the membrane shear elastic modulus (i.e., the resistance to elastic membrane deformation) was lower in the preterm infants than in the full-term neonates and lower in full-term infants than in adults. Thus, both whole cell and membrane deformability appear to be increased in preterm infants. Improved RBC deformability may aid in the maintenance of adequate blood circulation in preterm infants in spite of low vascular pressures.

Adult↗

Rigidity of elastomeric impression materials.

The rigidity of an impression material should be less than a certain value in order that excessive forces are not required when taking impressions of undercut areas. Conversely, the impression should be sufficiently rigid to withstand the weight of stone whilst pouring the cast without distortion. This paper describes a method of determining elastic modulus values at times as early as the setting time of the material. Results indicate that the elastic modulus values increase from the setting time to 30 min after the start of mix. Values are presented for a representative range of elastomeric impression materials.

Chemical Phenomena↗

Rheology of protein gels synthesized through a combined enzymatic and heat treatment method.

Whey protein gels prepared under acidic conditions (pH<4.6) remain largely unutilized because of their weak and brittle nature in contrast to the favorable elastic gels produced at neutral or basic conditions. However, such usage is important, as low pH food products are desirable due to their shelf stability and less stringent sterilization processes. In this study, we use a two-step process involving enzyme followed by heat treatment to produce whey protein gels at low pH (4.0). Dynamic rheological measurements reveal that the gel elastic modulus and yield stress increase substantially when heat treatment is supplemented with enzyme treatment. Both the elastic modulus and yield stress increase with increasing enzyme concentration or treatment time. In contrast, the dynamic yield strain decreases with enzyme concentration but increases with time of enzyme treatment. These results are explained in terms of the enzyme treatment time affecting the diffusion of the enzyme within the gel. This in turn leads to two types of gel microstructure at short and long enzyme treatment times, with the extent of enzyme diffusion modulating the structure at intermediate times.

Dose-Response Relationship, Drug↗

Composition and biomechanical properties of the bladder acellular matrix graft: comparative analysis in rat, pig and human.

OBJECTIVE: To compare the composition and mechanical properties of the newly developed bladder acellular matrix graft (BAMG) with the normal urinary bladder in rat, pig and human. MATERIALS AND METHODS: Rat, pig and human urinary bladders were harvested and divided into control and experimental groups. For the latter, BAMGs were prepared, and light and transmission electron microscopic studies performed. Strips from the normal bladders and the BAMGs (10 in each group) were tested under tension, and the ultimate tensile strength, maximum strain, and elastic modulus were determined from stress/strain curves. RESULTS: Both types I and III collagen, as well as elastic fibres, were observed as major components of the matrix scaffold. There were more collagen type I fibres in the rat than in the pig and human BAMGs, whereas the pig, and particularly the human, both showed higher levels of type III collagen and elastic fibres. These different matrix scaffold patterns were confirmed by electron microscopy. Results from biomechanical testing showed no significant differences for strength, strain or elastic modulus between BAMG and control bladder strips, except in the rat where the maximum strain values were significantly lower. CONCLUSION: There are variations in the acellular matrix structure with similar biomechanical properties between the BAMG and the normal urinary bladder in three different species. These results may underscore the potential of the BAMG. Furthermore, this in vitro model provides a suitable method to study the mechanical properties of the urinary bladder and may serve as a diagnostic tool for various investigations.

Animals↗

Comparison of pulse-wave velocity in different aortic regions in relation to the extent and severity of atherosclerosis between young and older Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits.

The present study was performed to investigate the effects of the development of atherosclerosis on foot-to-foot pulse-wave velocity (PWV) from the ascending aorta to different positions along the aorta in Kurosawa and Kusanagi-Hypercholesterolemic (KHC) rabbits aged 10-12 and 22-24 months old, in relation to the percentage fractional lesioned area (PFLA) in different aortic regions through which the pulse wave travels, as well as the rheological and pathohistological properties of the aortic wall. PWV, when measured in the KHC rabbit from the ascending aorta to each aortic position, showed the highest value on passage through the aortic arch, decreased with conduction to the distal thoracic aorta, reached the minimal value on passage to the distal thoracic aorta or to the middle abdominal aorta in the 10-12- and 22-24-month-old animals respectively, and increased gradually on conduction to the iliac artery. PWV at all aortic regions examined was significantly greater in the 22-24-month-old than in the 10-12-month-old KHC rabbits. PFLA, when measured in the aortic region from the ascending aorta to each aortic position, was maximal in the aortic arch and decreased gradually towards the peripheral aorta in both age groups. PFLA in the 22-24-month-old group was significantly greater than that in the 10-12-month-old group in all aortic regions examined. The atherosclerotic aortic wall showed a significantly lower elastic modulus in the young KHC rabbits compared with age-matched normal rabbits. A significantly higher elastic modulus was observed in the older KHC rabbits compared with that in the younger animals of both strains, associated with the progression of sclerosis. These findings indicate that the increase in PWV is due mainly to an increase with aging in the extent and severity of atherosclerosis in vessels in which the pulse wave travels.

Aging↗

Implementation of an optical method for the real-time determination of uniaxial strain and vessel mechanics.

The determination of the mechanical properties of vascular growth substrates has seen increasing interest in the bioengineering field. Mechanical features such as rupture strength, compliance characteristics, and viscoelastic properties of vascular grafts are important for their design and are indicative of their success in vivo. Thus a simple inexpensive measurement technique for these parameters would be useful. In this report we describe the implementation of an optical method for the measurement of vessel distention under a transluminal pressure gradient. It is based on the concept of laser light occlusion and allows for real time noncontact diameter measurements of hollow vessels < or = 2 cm. We demonstrate precise and reproducible measurements of diameter changes of less than 10 microm and, further, with the simultaneous determination of both strain and luminal pressure, were able to determine the elastic modulus of commercially available polymeric vessels. Comparison of the manufacturer specifications and our own measurement of the elastic modulus of these vessels, validate the effectiveness of our system. The advantages of this technique are its relative low cost, ease of implementation, high resolution, and flexibility stemming from its modular setup.

Animals↗

Hardness and yield strength of dentin from simulated nano-indentation tests.

The finite element method (FEM) is applied for studying the hardness (H) and yield strength (Y) of dentin subjected to a nano-indentation process. The nano-indentation experiments were simulated with the ABAQUS finite element software package. This test, performed with a spherical indenter, was simulated by axisymmetric finite element analysis. The load versus displacement was calculated during loading-unloading sequence for different elastic modulus (E) and yield strength. Hardness and maximum principal compressive and tensile stresses were plotted for different elastic modulus depending on yield strength. The dentin was assumed to be isotropic, homogenous and elasto-plastic. The theoretical results outlined in this study were compared with the experimental works reported in the literature and then hardness and yield strength of dentin was estimated.

Dentin↗

The prospects of estimating trabecular bone tissue properties from the combination of ultrasound, dual-energy X-ray absorptiometry, microcomputed tomography, and microfinite element analysis.

Osteoporosis commonly is assessed by bone quantity, using bone mineral density (BMD) measurements from dual-energy X-ray absorptiometry (DXA). However, such a measure gives neither information about the integrity of the trabecular architecture nor about the mechanical properties of the constituting trabeculae. We investigated the feasibility of deriving the elastic modulus of the trabeculae (the tissue modulus) from computer simulation of mechanical testing by microfinite element analysis (muFEA) in combination with measurements of ultrasound speed of sound (SOS) and BMD measurements. This approach was tested on 15 postmortem bovine bone cubes. The apparent elastic modulus of the specimens was estimated from SOS measurements in combination with BMD. Then the trabecular morphology was reconstructed using microcomputed tomography (muCT). From the reconstruction a mesh for muFEA was derived, used to simulate mechanical testing. The tissue modulus was found by correlating the apparent moduli of the specimens as assessed by ultrasound with the ones as determined with muFEA. A mean tissue modulus of 4.5 GPa (SD, 0.69) was found. When adjusting the muFEA-determined elastic moduli of the entire specimens with their calculated tissue modulus, an overall correlation of R2 = 96% with ultrasound-predicted values was obtained. We conclude that the apparent elastic stiffness characteristics as determined from ultrasound correlate linearly with those from muFEA. From both methods in combination, the elastic stiffness of the mineralized tissue can be determined as an estimator for mechanical tissue quality. This method can already be used for biopsy specimens, and potentially could be applicable in vivo as well, when clinical CT or magnetic resonance imaging (MRI) tools with adequate resolution reach the market. In this way, mechanical bone quality could be estimated more accurately in clinical practice.

Absorptiometry, Photon↗

Factors influencing left-ventricular stiffness.

The aim of the study was to investigate the relative contributions of geometrical and material factors to overall left-ventricular cavity stiffness. Left-ventricular cavity shapes were reconstructed using a computer and the variation of myocardial elastic modulus was calculated, by the finite element method, through the passive phase of diastole when rising volume coincided with rising pressure. Geometric data were obtained from biplane cineangiography, with micromanometer pressure measurements, for ten patients with left ventricular disease. Dimensional analysis was applied to the initial and derived data from which the influences of myocardial compliance, wall thickness-to-long dimension ratio, and aspect ratio (long-to-short axes) were determined. The ratio between the volume elasticity and the myocardial modulus of elasticity, the normalized stiffness ratio (NSR), is proposed as a useful index of left ventricular mechanical behaviour in diastole. The volume elasticity of the chamber is dependent not only upon the myocardium elastic modulus and the wall thickness ratio, but also on the shape of the chamber. Changes in the thickness/radius ratio of the ventricle have less effect upon its distention than those in the long dimension/radius ratio. The left ventricle becomes more spherical in shape through diastole and hence becomes stiffer by this geometric mechanism.

Cineangiography↗

Atomic force microscopy and nanoindentation characterization of human lamellar bone prepared by microtome sectioning and mechanical polishing technique.

Surface topography, microstructure, and micromechanical properties of human lamellar bone were characterized by atomic force microscopy and nanoindentation. The lamellar bone surfaces were prepared by two different methods: microtome sectioning and mechanical polishing. The lamellar bone surfaces prepared by mechanical polishing revealed that thin lamellae formed depressions approximately 200 nm deep, whereas the surfaces prepared by microtome sectioning were flat. Atomic force microscopy surface topographic images at higher magnification showed differences between thick and thin lamellae in polished samples, but these differences were less pronounced in microtomed samples. Roughness measurements confirmed that there was a significant difference between thick (21.0 nm) and thin lamellae (8.3 nm) in polished samples (p < 0.001). The difference in surface roughness between thick (13.9 nm) and thin lamellae (12.7 nm) in microtomed sample was statistically insignificant (p = 0.74). Higher elastic modulus values were observed for thick lamella in microtomed samples compared with that of thin lamellae, whereas measured elastic modulus differences between thick and thin lamellae in polished samples were found to be statistically insignificant.

Biomechanical Phenomena↗

Atomic force microscopy investigation of the dependence of cellular elastic moduli on glutaraldehyde fixation.

Summary The atomic force microscope (AFM) has provided nanoscale analyses of surfaces of cells that exhibit strong adhesive and cell spreading properties. However, it is frequently reported that prior fixation is required for reliable imaging of cells with lower adhesive properties. In the present study, the AFM is used to assess the effects of fixation by glutaraldehyde on the elastic modulus of a human rhabdomyosarcoma transfectant cell line RDX2C2. Our results show a sharp increase in the elastic modulus for even mild fixation (0.5% glutaraldehyde for 60 s), accompanied by a dramatic improvement in imaging reproducibility. An even larger increase is seen in NIH-3T3 mouse fibroblasts, although in that case fixation is not typically necessary for successful imaging. In addition, our results suggest that treatment with glutaraldehyde restricts the content of the resulting images to features nearer to the cell surface.

Animals↗

Influence of chemical structure on the fracture behaviour of dimethacrylate composite resins.

Previous studies have shown that the fracture resistance of dimethacrylate-based dental composite resins is enhanced by post-curing the matrix. Here, the influence of the chemical nature of the resin matrix is examined by a study of the fracture properties of composite resins formulated from 15 homologous dimethacrylate monomers and filled to 75 wt% with treated silica. The fracture toughness was determined via the double torsion technique and the elastic modulus and flexural strength were measured in flexure. The fracture energy calculated from the fracture toughness and elastic modulus, varied between 60 and 300 J/m2 while the fracture toughness ranged from 0.2 to 2.0 MN/m3/2 and the flexural strength varied from 17 to 111 MPa. The use of a blend of monomers was found to have a synergistic effect on the fracture resistance. Increasing the length of flexible spacer units (methylene or oxyethylene) between the methacrylate groups initially improved the fracture properties; however, beyond a certain length, these properties were impaired.

Acrylates↗