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Dimitrije Stamenović

Publications and source records attributed to Dimitrije Stamenović.

17 recordsLinked to original sources
PubMed2006

Contributions of the active and passive components of the cytoskeletal prestress to stiffening of airway smooth muscle cells.

Airway smooth muscle cells exhibit stiffening during contractile activation. This stiffening may be interpreted as a result of the stabilizing influence of the mechanical prestress stored within the cytoskeleton (CSK). However, in vivo, airway smooth muscle cells contract while simultaneously experiencing breathing-induced stretching. Excessive stretching of cells could cause actin-myosin crosslinks, and possibly other cytoskeletal filaments, to break, thereby leading to dissipation of the prestress and inhibition of further cell stiffening. The aim of this study is to investigate the stiffening behavior of individual human airway smooth muscle (HASM) cells exposed to a combination of substrate stretching, contractile activation and relaxation. We treated cultured HASM cells with either contractile (histamine) or relaxing (DBcAMP) pharmacological agonists and used magnetic cytometry technique to investigate the stiffening behavior of these cells during uniform substrate stretching (0-30%). Cells that were not treated, as well as those treated with histamine, exhibited increasing stiffening during stretching up to 20% of substrate strain, with additional stiffening becoming inhibited for substrate strains of 20-30%. In contrast, in cells treated with DBcAMP, stretching produced moderate but continuous stiffening with increasing substrate strain. These results indicate that both active and passive components of the prestress contribute to cell stiffening. We also observed that cells permeabilized with saponin exhibited stiffening at low levels (<10%) of substrate stretching, similar to non-permeabilized cells, but not at high levels (10-30%) of stretching, where stiffening was inhibited. These data suggest that at low levels of substrate strains the relative contributions of ion channel activation as well as actin and focal adhesion remodeling are less important for stiffening than passive distension of the CSK. Taken together, our results suggest that both the active and passive components of the cytoskeletal prestress contribute to the stiffening behavior of HASM cells under physiological conditions, but that at high levels of cellular distensions there is a possible tradeoff between these two components with the contribution from the passive component becoming increasingly more important.

Cells, Cultured↗
PubMed2006

Dynamics of prestressed semiflexible polymer chains as a model of cell rheology.

We report on a model of a prestressed nonlinear semiflexible polymer chain that links thermally driven dynamics to the creep behavior of living cells. Numerical simulations show that the chain's creep follows a power law with an exponent that decreases with increasing prestress. This is related to the propagation of free energy through the chain in response to stretching, where the propagation speed is regulated by the prestress via the chain's nonlinear elasticity. These results indicate that the main aspects of cell rheology are consistent with the dynamics of single polymer chains under tension.

Actins↗
PubMed2006

A mathematical model of cell reorientation in response to substrate stretching.

It is well documented that in response to substrate stretching adhering cells alter their orientation. Generally, the cells reorient away from the direction of the maximum substrate strain, depending upon the magnitude of the substrate strain and the state of cell contractility. Theoretical models from the literature can describe only some aspects of this phenomenon. In the present study, we developed a more comprehensive mathematical model of cell reorientation than the current models. Using the framework of theory of non-linear elasticity, we found that the problem of cell reorientation was a stability problem, with the global (Maxwell's) criterion for stability. For the case of uniaxial substrate stretching, we showed that cells would orient away from the direction of substrate strain such that the angle between the cell long axis and the direction of the substrate strain would increase with increasing magnitude of the strain. We also showed that at a given substrate strain this angle would be greater in cells having greater contractile strain. These results are consistent with experimental observations reported in the literature.

Animals↗
PubMed2005

Effects of cytoskeletal prestress on cell rheological behavior.

Normal tissue development requires that cells alter their mechanical behavior in different microenvironments to carry out their diverse functions. During cell spreading, migration, invasion and mitosis, cells exhibit a high degree of deformability, exhibiting almost a fluid-like behavior, whereas within quiescent differentiated tissues, cells must behave like an elastic solid to maintain their structural integrity in the face of an applied mechanical stress. A growing body of experimental evidence suggests that rheological properties of adherent cells depend on pre-existing tensional stress ("prestress") borne by the cytoskeleton. This prestress results from the action of tensional forces borne by actin microfilaments, transmitted over intermediate filaments and resisted by both extracellular matrix adhesions and internal microtubules. Observations that the prestress influences mechanical properties of the cell are intimately related to the cellular tensegrity model. This model depicts the cytoskeleton as an interconnected network of cables that carry pre-existing tension that is balanced by compression-bearing struts and by anchoring forces of the substrate. This paper offers a brief survey of the basic concept of cellular tensegrity model, comparison of model predictions with experimental data obtained from rheological measurements on living cells, and comparison with other models that have been used in studies of rheology of cells.

Cells↗
PubMed2005

Microtubules may harden or soften cells, depending of the extent of cell distension.

Experimental data show that disruption of microtubules causes cells to either become stiffer or softer. Current understanding of these behaviors is based on several different mechanisms, each of which can account for only stiffening or softening. In this study we offer a model that can explain both these features. The model is based on the cellular tensegrity idea. Key premises of the model are that cell shape stability is secured through pre-existing mechanical stress (prestress) borne by the actin cytoskeletal network, and that this prestress is partly balanced by cytoskeletal microtubules and partly by the extracellular matrix. Thus, disturbance of this balance would affect cell deformability. The model predicts that disruption of microtubules causes an increase or a decrease in cell stiffness, depending on the extent to which microtubules participate in balancing the prestress which, in turn, depends on the extent of cell spreading. In highly spread cells microtubules have a minor and negative contribution to cell stiffness, whereas in less spread cells their contribution is positive and substantial. Since in their natural habitat cells seldom exhibit highly spread forms, the above results suggest that the contribution of microtubules to cell deformability cannot be overlooked.

Animals↗
PubMed2005

Contractile torque as a steering mechanism for orientation of adherent cells.

It is well established that adherent cells change their orientation in response to non-uniform substrate stretching. Most observations indicate that cells orient away from the direction of the maximal substrate strain, whereas in some cases cells also align with the direction of the maximal strain. Previous studies suggest that orientation and steering of the cell may be closely tied to cytoskeletal contractile stress but they could not explain the mechanisms that direct cell reorientation. This led us to develop a simple, mechanistic theoretical model that could predict a direction of cell orientation in response to mechanical nonuniformities of the substrate. The model leads to a simple physical mechanism--namely the contractile torque--that directs the cell toward a new orientation in response to anisotropic substrate stretching or substrate material anisotropy. A direction of the torque is determined by a dependence of the contractile stress on substrate strain. Model predictions are tested in the case of simple elongation of the substrate and found to be consistent with experimental data from the literature.

Cell Adhesion↗
PubMed2004

Distending stress of the cytoskeleton is a key determinant of cell rheological behavior.

One fundamental question in cell biology is what determines rheological properties of living cells. If the cytoskeletal distending stress is a key determinant of cell rheology, then modulating this stress by cell stretching should have a major effect on cell rheological properties. If not, then other mechanisms must play a major role. We developed a stretchable cell culture device that could rapidly stretch cells and thus generate passive mechanical stress within the cytoskeleton. This device was placed inside a magnetic cytometry system to measure the effect of stretching on rheological properties of cultured human airway smooth muscle cells. A gradual increase in cell distension caused a systematic increase in cell dynamic stiffness in a manner which was consistent with earlier observations where the active component of the distending stress was modulated pharmacologically. These findings provide strong evidence that the cytoskeletal distending stress is a key determinant of cell rheological properties.

Cell Culture Techniques↗
PubMed2004

A computational tensegrity model predicts dynamic rheological behaviors in living cells.

Rheological properties of living cells play a key role in the control of cell shape, growth, movement, and contractility, yet little is known about how these properties are governed. Past approaches to understanding cell mechanics focused on the contributions of membranes, the viscous cytoplasm, and the individual filamentous biopolymers that are found within the cytoskeleton. In contrast, recent work has revealed that the dynamic mechanical behavior of cells depends on generic system properties, rather than on a single molecular property of the cell. In this paper, we show that a mathematical model of cell mechanics that depicts the intracellular cytoskeleton as a tensegrity structure composed of a prestressed network of interconnected microfilaments, microtubules, and intermediate filaments, and that has previously explained static cellular properties, also can predict fundamental dynamic behaviors of living cells.

Actin Cytoskeleton↗
PubMed2003

A prestressed cable network model of the adherent cell cytoskeleton.

A prestressed cable network is used to model the deformability of the adherent cell actin cytoskeleton. The overall and microstructural model geometries and cable mechanical properties were assigned values based on observations from living cells and mechanical measurements on isolated actin filaments, respectively. The models were deformed to mimic cell poking (CP), magnetic twisting cytometry (MTC) and magnetic bead microrheometry (MBM) measurements on living adherent cells. The models qualitatively and quantitatively captured the fibroblast cell response to the deformation imposed by CP while exhibiting only some qualitative features of the cell response to MTC and MBM. The model for CP revealed that the tensed peripheral actin filaments provide the key resistance to indentation. The actin filament tension that provides mechanical integrity to the network was estimated at approximately 158 pN, and the nonlinear mechanical response during CP originates from filament kinematics. The MTC and MBM simulations revealed that the model is incomplete, however, these simulations show cable tension as a key determinant of the model response.

Actin Cytoskeleton↗
PubMed2003

Fractional derivatives embody essential features of cell rheological behavior.

Mechanical moduli of cultured airway smooth muscle cells (Fabry, B., et al. Phys. Rev. Lett. 87:148102, 2001) reveal that the frequency dependence of cell rheological behavior conforms to a weak power-law relationship over a wide range of frequency (10(-2)-10(3) Hz). Such a behavior cannot be accounted for by standard viscoelastic models characterized by a discrete number of time constants that have been commonly used in previous studies of cell viscoelasticity. Fractional calculus, by contrast, provides a natural framework for describing weak power-law relationships and requires no assumptions about the type of material, the time constant distribution or the time/frequency interval in which rheological observations are made. In this study, we developed a rheological model of the cell using methods of fractional calculus. We used a least-squares technique to fit the model to data previously obtained from measurements on airway smooth muscle cells. The fit provided an excellent correspondence to the data, and the estimated values of model parameters were physically plausible. The model leads to a novel and unexpected empirical link between dynamic viscoelastic behavior of the cytoskeleton and the static contractile stress that it bears.

Animals↗
PubMed2003

Experimental tests of the cellular tensegrity hypothesis.

The tensegrity model depicts the cytoskeleton (CSK) as a prestressed network of interconnected filaments. The prestress is generated by the CSK contractile apparatus and is partly balanced by traction at the cell-substrate interface and partly by CSK internal compression elements such as microtubules (MTs). A key feature of tensegrity is that the shear modulus (G) must increase in proportion with the prestress. Here we have tested that prediction as well as the idea that compression of MTs balance a portion of the cell prestress. Airway smooth muscle cells were studied. Traction microscopy was used to calculate traction. Because traction must be balanced by the stress within the cell, the prestress could be computed. Cell G was measured by oscillatory magnetic cytometry. The prestress was modulated using graded concentrations of contracting (histamine) or relaxing (isoproterenol) agonists and by disrupting MTs by colchicine. It was found that G increased in proportion with the prestress and that compression of MTs balanced a significant, but a relatively small fraction of the prestress. Taken together, these results do not disprove other models of cell deformability, nor they prove tensegrity. However, they do support a priori predictions of tensegrity. As such, it may not be necessary to invoke more complex mechanisms to explain these central features of cell deformability.

Cell Culture Techniques↗
PubMed2002

Physiology: Dynamic instabilities in the inflating lung.

In lung diseases such as asthma, expiratory flow becomes limited, airways can collapse and the vital exchange of gases is compromised. Here we model the inflation of collapsed regions of the lung during inspiration in terms of avalanches propagating through a bifurcating network of airways, and find that the accompanying cascade of dynamic pressure instabilities -- avalanche 'shocks' -- manifests as negative elastic resistance of the lung. Our analysis of this apparent thermodynamic paradox provides a better understanding of aeration in the deep regions of the lung, which may find application in medical conditions in which gas exchange is impaired.

Animals↗
PubMed2002

Cell prestress. I. Stiffness and prestress are closely associated in adherent contractile cells.

The tensegrity hypothesis holds that the cytoskeleton is a structure whose shape is stabilized predominantly by the tensile stresses borne by filamentous structures. Accordingly, cell stiffness must increase in proportion with the level of the tensile stress, which is called the prestress. Here we have tested that prediction in adherent human airway smooth muscle (HASM) cells. Traction microscopy was used to measure the distribution of contractile stresses arising at the interface between each cell and its substrate; this distribution is called the traction field. Because the traction field must be balanced by tensile stresses within the cell body, the prestress could be computed. Cell stiffness (G) was measured by oscillatory magnetic twisting cytometry. As the contractile state of the cell was modulated with graded concentrations of relaxing or contracting agonists (isoproterenol or histamine, respectively), the mean prestress ((t)) ranged from 350 to 1,900 Pa. Over that range, cell stiffness increased linearly with the prestress: G (Pa) = 0.18(t) + 92. While this association does not necessarily preclude other interpretations, it is the hallmark of systems that secure shape stability mainly through the prestress. Regardless of mechanism, these data establish a strong association between stiffness of HASM cells and the level of tensile stress within the cytoskeleton.

Acrylic Resins↗
PubMed2002

Cell prestress. II. Contribution of microtubules.

The tensegrity model hypothesizes that cytoskeleton-based microtubules (MTs) carry compression as they balance a portion of cell contractile stress. To test this hypothesis, we used traction force microscopy to measure traction at the interface of adhering human airway smooth muscle cells and a flexible polyacrylamide gel substrate. The prediction is that if MTs balance a portion of contractile stress, then, upon their disruption, the portion of stress balanced by MTs would shift to the substrate, thereby causing an increase in traction. Measurements were done first in maximally activated cells (10 microM histamine) and then again after MTs had been disrupted (1 microM colchicine). We found that after disruption of MTs, traction increased on average by approximately 13%. Because in activated cells colchicine induced neither an increase in intracellular Ca(2+) nor an increase in myosin light chain phosphorylation as shown previously, we concluded that the observed increase in traction was a result of load shift from MTs to the substrate. In addition, energy stored in the flexible substrate was calculated as work done by traction on the deformation of the substrate. This result was then utilized in an energetic analysis. We assumed that cytoskeleton-based MTs are slender elastic rods supported laterally by intermediate filaments and that MTs buckle as the cell contracts. Using the post-buckling equilibrium theory of Euler struts, we found that energy stored during buckling of MTs was quantitatively consistent with the measured increase in substrate energy after disruption of MTs. This is further evidence supporting the idea that MTs are intracellular compression-bearing elements.

Acrylic Resins↗
PubMed2002

Effect of surface tension on alveolar surface area.

At fixed lung volume (VL), alterations in surface tension change alveolar surface area (S) and lung recoil (PL). Wilson (26), using data from fixed lungs (1, 9), quantified the isovolume change in S with PL. We reexamined this question in fresh excised rabbit lungs, with two important differences. First, we measured fractional changes in S by using diffuse light scattering, avoiding the potential upset of the balance of tissue and surface forces during fixation. Second, we altered surface tension by ventilating the lungs with nebulized polydimethylsiloxane, with much less residual fluid compared with lavage. We found that S decreased at low and mid VL (treatment surface tension > control) by about half of Wilson's estimates and was nearly unaffected by treatment at high VL. This suggests that with increased surface tension there is 1) greater septal retraction in lungs fixed by vascular perfusion compared with unfixed lungs and 2) a greater increase in PL and less loss of S than would have been predicted.

Animals↗
PubMed2002

Effect of the cytoskeletal prestress on the mechanical impedance of cultured airway smooth muscle cells.

We investigated the effect of the cytoskeletal prestress (P) on the elastic and frictional properties of cultured human airway smooth muscle cells during oscillatory loading; P is preexisting tensile stress in the actin cytoskeleton generated by the cell contractile apparatus. We oscillated (0.1 Hz, 6 Pa peak to peak) small ferromagnetic beads bound to integrin receptors and computed the storage (elastic) modulus (G') and the loss (frictional) modulus (G") from the applied torque and the corresponding bead rotation. All measurements were done at baseline and after cells were treated with graded doses of either histamine (0.1, 1, 10 microM) or isoproterenol (0.01, 0.1, 1, 10 microM). Values for P for these concentrations were taken from a previous study (Wang et al., Am J Physiol Cell Physiol, in press). It was found that G' and G", as well as P, increased/decreased with increasing doses of histamine/isoproterenol. Both G' and G" exhibited linear dependences on P: G'(Pa) = 0.20P + 82 and G"(Pa) = 0.05P + 32. The dependence of G' on P is consistent with our previous findings and with the behavior of stress-supported structures. The dependence of G" on P is a novel finding. It could be attributed to a variety of mechanisms. Some of those mechanisms are discussed in detail. We concluded that, in addition to the central mechanisms by which stress-supported structures develop mechanical stresses, other mechanisms might need to be invoked to fully explain the observed dependence of the cell mechanical properties on the state of cell contractility.

Actins↗