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Mechanical stimuli are fundamental in the development of organs and tissues, their growth, regeneration or disease. They influence the biochemical signals produced by the cells, and, consequently, the development and spreading of a disease. Moreover, tumour cells are usually characterized by a decrease in the cell mechanical properties that may be directly linked to their metastatic potential. Thus, recently, the experimental and computational study of cell biomechanics is facing a growing interest. Various experimental approaches have been implemented to describe the passive response of cells; however, cell variability and complex experimental procedures may affect the obtained mechanical properties. For this reason, in-silico computational models have been developed through the years, to overcome such limitations, while proposing valuable tools to understand cell mechanical behaviour. This being the case, we propose a combined continuous-tensegrity finite element (FE) model to analyse the mechanical response of a cell and its subcomponents, observing how every part contributes to the overall mechanical behaviour. We modelled both Atomic Force Microscopy (AFM) indentation and micropipette aspiration techniques, as common mechanical tests for cells and elucidated also the role of cell cytoplasm and cytoskeleton in the global cell mechanical response. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Cellular mechanical characteristics represent cell ability to produce tissue-specific metabolites. Therefore, to achieve effective cell therapy, a better understanding of the effects of chemo-mechanical stimuli on the mechanical properties of in vitro-treated cells is essential. Herein, we investigated the effects of uniaxial strain on the mechanical properties of mesenchymal stem cells (MSCs) upon transforming growth factor beta 1 (TGF-β1) stimulation. The MSCs were categorized into control and test groups. In one test group, the MSCs were treated by TGF-β1 for 6 d, and in the other, they were additionally subjected to 1-d uniaxial strain on day 2. The cell mechanical properties and smooth muscle (SM) gene expression were assessed on days 2, 4, and 6. During the entire experiment, the MSCs treated by TGF-β1 ± uniaxial strain were induced to differentiate into SM-like cells by significantly upregulation of α-actin, SM22α, and h1-calponin in respect to the control samples. When the MSCs were treated with TGF-β1 alone, their stiffness and viscosity decreased significantly on day 2 and then increased by increase in culture time. When the cells were subjected to 1-d uniaxial strain upon TGF-β1 stimulation, their stiffness and viscosity significantly increased on days 2 and 4 and then decreased on day 6 to a level comparable to that of TGF-β1 group. Different paths were noticeable among the treated samples to reach nearly similar states on day 6. It seems that uniaxial strain activates mechanobiological cascades by which cellular mechanical behavior can be regulated after its removal. However, these effects are transient and would diminish over time. The findings may be helpful in the chemo-mechanical regulation of MSCs. [ABSTRACT FROM AUTHOR]

In this study, an aspiration system for elastic spheres was developed to verify the approximate elastic formula for the aspirated length of a single solid-like cell undergoing micropipette aspiration (MPA), which was obtained in our previous study by theoretical analysis and numerical simulation. Using this system, foam silicone rubber spheres with different diameters and mechanical properties were aspirated in a manner similar to the MPA of single cells. Comparisons between the approximate elastic formula and aspiration experiments of spheres indicated that the predictions of the formula agreed with the experimental results. Additionally, combined with the MPA data of rabbit chondrocytes, differences in terms of the elastic parameters derived from the half-space model, incompressible sphere model, and compressible sphere model were explored. The results demonstrated that the parameter ξ (ξ = R/a, where R is the radius of the cell and a is the inner radius of the micropipette) and Poisson’s ratio significantly influenced the determination of the elastic modulus and bulk modulus of the cell. This work developed for the first time an aspiration system of elastic spheres to study the elastic responses of the MPA of a single cell and provided new evidence supporting the use of the approximate elastic formula to determine cellular elastic parameters from the MPA data. [ABSTRACT FROM AUTHOR]

Defining the characteristics of cancer stem cells (CSC) has become an important subject in cancer research during the past decade. Although molecular surface expression levels have been used for CSC recognition, the clinical and prognostic impacts of these markers have remained a controversial issue. The finding that cancerous cells are considerably more deformable than normal ones provides the motivation for the hypothesis that the mechanical properties can be used as biomarkers to distinguish between stem-like and non-stem-like cancer cells. In this study, using micropipette aspiration (MA) and intracellular particle tracking (IPT) microrheology, measurements of the whole-cell and local viscoelasticity were made on four breast cancer cell lines with different CSC phenotypes based on their surface markers. Stem-like Hs578T and MDA-MB-231 cell lines were found to be the most deformable, while the non-stem-like MDA-MB-468 line was the least deformable. The non-stem-like BT-20 cell line showed an intermediate deformability. The enhanced deformability for stem-like cells was consistent with the observed lower and more dispersed F-actin content for the stem-like cells. Therefore, the cytoskeleton-related differences in the rheological properties of cancer cells can be a potential biomarker for CSC and eventually lead to novel cancer diagnostic and therapeutic methods. [ABSTRACT FROM AUTHOR]

The cellular structures and mechanical properties of human mesenchymal stem cells (hMSCs) vary significantly during culture and with differentiation. Previously, studies to measure mechanics have provided divergent results using different quantitative parameters and mechanical models of deformation. Here, we examine hMSCs prepared for clinical use and subject them to mechanical testing conducive to the relevant deformability associated with clinical injection procedures. Micropipette aspiration of hMSCs shows deformation as a viscoelastic fluid, with little variation from cell to cell within a population. After two passages, hMSCs deform as viscoelastic solids. Further, for clinical applicability during stem cell migration in vivo, we investigated the ability of hMSCs to invade into micropillar arrays of increasing confinement from 12 to 8 μm spacing between adjacent micropillars. We find that hMSC samples with reduced deformability and cells that are more solid-like with passage are more easily able to enter the micropillar arrays. Increased cell fluidity is an advantage for injection procedures and optimization of cell selection based on mechanical properties may enhance efficacy of injected hMSC populations. However, the ability to invade and migrate within tight interstitial spaces appears to be increased with a more solidified cytoskeleton, likely from increased force generation and contractility. Thus, there may be a balance between optimal injection survival and in situ tissue invasion. [ABSTRACT FROM AUTHOR]

Cell adhesion plays an important role in cell physiology. A better understanding of this process could facilitate many clinical therapies. In this study, Rat bone marrow-derived mesenchymal stem cells (rBMSCs) were cultured on glass substrate, and the morphology and adhesion strength were characterized. The cell morphology was defined as spherical, adhesive, and spreading. The adhesion strengths of the different morphologies exhibited different distribution patterns. The spherical cells exhibited low adhesion strength; the adhesive cells exhibited rapidly increasing adhesion strength while their diameters remained relatively constant. The adhesion strength increased with the cell diameter in the spreading cells. These findings suggest that adhesion strength can be quickly assessed by examining the cell morphology. [ABSTRACT FROM AUTHOR]

The interaction of leukocytes with surface bound ligands can be limited by the location of the molecules relative to the surface topology of the cell. In this report, we examine the dynamic response of neutrophils to IL-8-fractalkine chimera immobilized on bead surfaces, taking into account changes in receptor occupancy resulting from changes in surface topography. As a readout for receptor signaling, we observe the dynamics of calcium release in neutrophils following contact with the IL-8 coated surface. After a delay that depended on the initial area of contact and the surface density of IL-8, the cell began to phagocytose the IL-8 coated bead. This appeared to be a pre-requisite for release of calcium, which typically followed shortly after the initiation of phagocytosis. In separate experiments, effective kinetic coefficients for the formation of bonds between immobilized IL-8 and receptors on the cell surface were determined. Using these coefficients, we were able to estimate the number of bound receptors in the nascent contact zone. Kinetic modeling of the signaling response predicted that cell spreading and a concomitant increase in the density of occupied receptors would be required for the experimentally observed calcium dynamics. Postulating that there is an increase in receptor occupancy resulting from smoothing of the cell surface as it is stretched over the bead enabled us to obtain model predictions consistent with experimental observations. This study reveals the likely importance of membrane microtopology as a rate-limiting property and potential means of regulation of cell responses stimulated by two-dimensional surface interactions. [ABSTRACT FROM AUTHOR]

Aortopathy is characterized by vascular smooth muscle cell (VSMC) abnormalities and elastic fiber fragmentation. Elastin insufficient ( Eln) mice demonstrate latent aortopathy similar to human disease. We hypothesized that aortopathy manifests primarily in the aorto-pulmonary septal (APS) side of the thoracic aorta due to asymmetric cardiac neural crest (CNC) distribution. Anatomic (aortic root vs. ascending aorta) and molecular (APS vs. non-APS) regions of proximal aorta tissue were examined in adult and aged wild type (WT) and mutant ( Eln) mice. CNC, VSMCs, elastic fiber architecture, proteoglycan expression, morphometrics and biomechanical properties were examined using histology, 3D reconstruction, micropipette aspiration and in vivo magnetic resonance imaging (MRI). In the APS side of Eln aorta, Sonic Hedgehog (SHH) is decreased while SM22 is increased. Elastic fiber architecture abnormalities are present in the Eln aortic root and APS ascending aorta, and biglycan is increased in the aortic root while aggrecan is increased in the APS aorta. The Eln ascending aorta is stiffer than the aortic root, the APS side is thicker and stiffer than the non-APS side, and significant differences in the individual aortic root sinuses are observed. Asymmetric structure-function abnormalities implicate regional CNC dysregulation in the development and progression of aortopathy. [ABSTRACT FROM AUTHOR]

Human mesenchymal stem cells (hMSCs) are multipotent cells appropriate for a variety of tissue engineering and cell therapy applications. Mechanical properties of hMSCs during differentiation are associated with their particular metabolic activity and regulate cell function due to alternations in cytoskeleton and structural elements. The objective of this study is to evaluate elastic and viscoelastic properties of hMSCs during long term cultivation in control and transforming growth factor-β treatment groups using micropipette aspiration technique. The mean Young's modulus ( E) of the control samples remained nearly unchanged during 6 days of cultivation, but that of the test samples showed an initial reduction compared to its relevant control sample after 2 days of treatment by biological growth factor, followed by a significant rise after 4 and 6 days. The viscoelastic creep tests showed that both instantaneous and equilibrium moduli significantly increased with the treatment time and reached to maximum values of 622.9 ± 114.2 and 144.3 ± 11.6 Pa at the sixth day, respectively, while increase in apparent viscosity was not statistically significant. Such change of mechanical properties of hMSCs during specific lineage commitment contributes to regenerative medicine as well as stem-cell-based therapy in which biophysical signals regulate stem cell fate. [ABSTRACT FROM AUTHOR]

Micropipette aspiration (MA) has been widely used to measure the biomechanical properties of cells and biomaterials. To estimate material parameters from MA experimental data, analytical half-space models and inverse finite element (FE) analyses are typically used. The half-space model is easy to implement but cannot account for nonlinear material properties and complex geometrical boundary conditions that are inherent to MA. Inverse FE approaches can account for geometrical and material nonlinearities, but their implementation is resource-intensive and not widely available. Here, by making analogy between an analytical uniaxial tension model and a FE model of MA, we proposed an easily implementable and accurate method to estimate the material parameters of tissues tested by MA. We first adopted a strain invariant-based isotropic exponential constitutive model and implemented it in both the analytical uniaxial tension model and the FE model. The two models were fit to experimental data generated by MA of porcine aortic valve tissue (45 spots on four leaflets) to estimate material parameters. We found no significant differences between the effective moduli estimated by the two models ( $$p > 0.39$$), with the effective moduli estimated by the uniaxial tension model correlating significantly with those estimated by the FE model ( $$p < 0.001; R^{2}= 0.96$$) with a linear regression slope that was not different than unity ( $$p = 0.38$$). Thus, the analytical uniaxial tension model, which avoids solving resource-intensive numerical problems, is as accurate as the FE model in estimating the effective modulus of valve tissue tested by MA. [ABSTRACT FROM AUTHOR]

Micropipette aspiration (MA) is widely applied in cell mechanics, however, at small deformations a common model corresponding to the MA is the half-space model wherein the finite cell size and cell compressibility are neglected. This study extends the half-space model by accounting for the influence of cell geometry and compressibility (sphere model). Using a finite element analysis of cell aspiration into a micropipette, an elastic approximation formula of the aspirated length was derived for the sphere model. The approximation formula includes the geometry parameter ξ of the sphere model ( ξ= R/ a, R is the radius of the cell, and a is the inner radius of the micropipette) and the Poisson's ratio v of the cell. The results indicate that the parameter ξ and Poisson's ratio v markedly affect the aspirated length, particularly for small ξ and v. When ξ→∞ and v→0.5, the approximation formula tends to the analytical solution for the half-space model. In the incompressible case ( v = 0.5), within the general experimental range ( ξ varying from 2 to 4), the difference between the analytical solution and the approximate one is significant, and is up to 29% of the approximation solution when ξ = 2. Additionally, parametere was introduced to evaluate the error of elastic moduli between the half-space model and sphere model. Based on the approximation formula, the ξ thresholds, beyond which e becomes larger than 10% and 20%, were derived. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate effects of cyclic stretch on the mechanical properties of Endothelial cells (ECs). Human Umbilical Vein Endothelial Cells were cultured and exposed to uniaxial cyclic stretch with two amplitude (10 % and 20 %) and different time duration (2, 4, 6 and 8 h). Micropipette aspiration technique coupled with the generalized Maxwell model was used to evaluate the mechanical properties of the ECs. Effects of cyclic stretch on the cell cytoskeleton were analyzed by actin filament staining. Results confirmed viscoelastic behavior of ECs in the micropipette aspiration experiment. The present results confirmed that increased stretch amplitude and duration led to lower deformation and further stiffening of ECs. Actin fibers were shown to align and bundle in response to the cyclic stretch. The results were compared statistically among test and control groups and it was concluded that cyclic loading causes significant alteration in mechanical properties of ECs due to remodeling of cell cytoskeleton. [ABSTRACT FROM AUTHOR]

This paper presents a system for mechanically characterizing single cells using automated micropipette aspiration. Using vision-based control and position control, the system controls a micromanipulator, a motorized translation stage, and a custom-built pressure system to position a micropipette (4 μm opening) to approach a cell, form a seal, and aspirate the cell into the micropipette for quantifying the cell's elastic and viscoelastic parameters as well as viscosity. Image processing algorithms were developed to provide controllers with real-time visual feedback and to accurately measure cell deformation behavior on line. Experiments on both solid-like and liquid-like cells demonstrated that the system is capable of efficiently performing single-cell micropipette aspiration and has low operator skill requirements. [ABSTRACT FROM AUTHOR]

Background: The study aims to determine the changes in the biomechanical properties of the anterior and extreme posterior portions of experimental near-sighted eyes by examining the mechanical behavior of guinea pig scleral desmocytes, thus finding a new approach to the pathogenesis of myopia and their corresponding therapies. Methods: Guinea pigs (2 weeks old) were numbered and assigned into three groups (A, B, and C) with ten guinea pigs each. Concave lens-induced myopic (LIM) animal models were prepared via the out-of-focus method. The other eye in the same guinea pig served as the self-control (SC) group. After modeling groups A, B, and C for 6, 15, and 30 days respectively, the lenses from the guinea pigs in the experimental group were removed. The scleral fibroblasts in each group were cultured, and passaged twice in vitro. The micropipette aspiration technique coupled with a viscoelastic solid model was utilized to investigate the viscoelastic properties of the scleral fibroblasts in normal and myopic guinea pigs. The mechanical behavior of the scleral desmocytes of the LIM and SC groups were compared. Results: The mechanical behavior of the scleral desmocytes was compared between the LIM and SC groups. The Young's modulus at equilibrium and the apparent cellular viscosity of the anterior portion of the sclera in the LIM group at 6 days and 15 days after myopic induction were not significantly different from that of the SC group ( P < 0.05). However, the results for the anterior portions of the sclera in the LIM group at 30 days were significantly higher than those of the LIM group at 6 and 15 days, as well as those in the SC group ( P < 0.05). The Young's modulus at equilibrium and the apparent cellular viscosity of the extreme posterior portions of the sclera in the LIM group at 6 days after myopic induction not significantly from those of the SC group ( P < 0.05). However, the results for the extreme posterior portions of the sclera in the LIM group after 15 days and 30 days were significantly higher than those in the LIM group at 6 days and the SC group ( P < 0.05). Conclusions: The Young's modulus at equilibrium or apparent cellular viscosity of all the anterior portions of the sclera in the LIM group were longer than those in the SC group at 30 days after the induction, and the results for all the extreme posterior portions of the LIM group were larger than those of the SC group on the 15th and 30th day. Therefore, the Young's modulus and apparent viscosity of the anterior and extreme posterior portions of the sclera changed on the 15th and 30th day after induction respectively. [ABSTRACT FROM AUTHOR]

The unfertilized oocyte is surrounded by a spherical layer called the zona pellucida (ZP). The physical hardness of this layer plays a crucial role in fertilization and it is largely unknown because of the lack of appropriate measuring and modeling methods. Recently, considerable biomedical attentions have concentrated on determination of the mechanical properties of oocytes as a single cell. In order to investigate the biophysical characteristics of mammalian oocytes, a change in the elasticity of human ZP has been quantitatively evaluated before and after fertilization. Young's modulus of ZP of metaphase-II (MII) and pronuclear (PN) stages have been estimated using two different protocols of the micropipette aspiration, step-by-step and continuous increase in pressure, in combination with proportional theoretical models. Experimental results clearly demonstrated that after fertilization the mean Young's modulus of the ZP calculated from the step-by-step aspiration test (MII: 7.34 ± 1.36 kPa vs PN: 13.18 ± 1.17 kPa.) and continuous aspiration test (MII: 2.41 ± 0.75 kPa vs. PN: 4.43 ± 1.66 kPa) significantly increased, ( p < 0.05). Mathematical Evaluation of the results shows that although the results of the two methods are different but both confirm that the hardening of ZP will increase following fertilization. As can be seen, different experimental methods can influence the choice of the models and this in turn will lead the mechanical properties to be found. [ABSTRACT FROM AUTHOR]

Membrane tethers or nanotubes play a critical role in a variety of cellular and subcellular processes such as leukocyte rolling and intercellular mass transport. The current constitutive equations that describe the relationship between the pulling force and the tether velocity during tether extraction have serious limitations. In this article, we propose a new phenomenological constitutive equation that captures all known characteristics of nanotube formation, including nonlinearity, nonzero threshold force, and possible negative tether velocity. We used tether extraction from endothelial cells as a prototype to illustrate how to obtain the material constants in the constitutive equation. With the micropipette aspiration technique, we measured tether pulling forces at both positive and negative tether velocities. We also determined the threshold force of 55 pN experimentally for the first time. This new constitutive equation unites two established ones and provides us a unified platform to better understand not only the physiological role of tether extraction during leukocyte rolling and intercellular or intracellular transport, but also the physics of membrane tether growth or retraction. [ABSTRACT FROM AUTHOR]

The natural biconcave shape of red blood cells (RBC) may be altered by injury or environmental conditions into a spiculated form (echinocyte). An analysis is presented of the effect of such a transformation on the resistance of RBC to entry into capillary sized cylindrical tubes. The analysis accounts for the elasticity of the membrane skeleton in dilation and shear, and the local and nonlocal resistance of the bilayer to bending, the latter corresponding to different area strains in the two leaflets of the bilayer. The shape transformation is assumed to be driven by the equilibrium area difference (ΔA[sub 0], the difference between the equilibrium areas of the bilayer leaflets), which also affects the energy of deformation. The cell shape is approximated by a parametric model. Shape parameters, skeleton shear deformation, and the skeleton density of deformed membrane relative to the skeleton density of undeformed membrane are obtained by minimization of the corresponding thermodynamic potential. Experimentally, ΔA[sub 0] is modified and the corresponding discocyte–echinocyte shape transition obtained by high-pressure aspiration into a narrow pipette, and the deformability of the resulting echinocyte is examined by whole cell aspiration into a larger pipette. We conclude that the deformability of the echinocyte can be accounted for by the mechanical behavior of the normal RBC membrane, where the equilibrium area difference ΔA[sub 0] is modified. [ABSTRACT FROM AUTHOR]