Felix A. Blyakhman

A correlation between mechanical and electrical properties of the synthetic hydrogel chosen as an experimental model of cytoskeleton

The correlation between the electrochemical (Donnan) potential and volume swelling was studied for synthetic polyelectrolyte hydrogels considered as models of cytoskeleton gel-forming biopolymers. Hydrogels involving polyacrylic and polymethacrylic acids with varying network density were synthesized by a radical polymerization in aqueous solution. Electrical charge was introduced into the gel network by partial neutralization of monomer acids with several alkali and alkali earth (hydr)oxides. The electrochemical (Donnan) potential of synthetic gels was determined using conventional microelectrode tools for cell potential determination. It was demonstrated that the negative electrical potential of many anionic gels with various charges and network densities decreased with the decrease of equilibrium swelling, i.e., with the decrease in water content in the gel. It was shown that a drastic phase transition in the gel structure from a swollen to a compressed state induced by K+/Ca2+ exchange is accompanied by an analogous decrease in the absolute Donnan potential of the gels. A kinetic study demonstrated that the gel volume changed ahead of its electrical potential. This suggests that the volume phase transition in gel is the main cause of the electrical response. A similarity between the swelling/compression transition in synthetic gels and the volume changes in the cytoskeleton in the vicinity of the cell membrane was demonstrated. Based on the universal analogy between the properties of synthetic and natural polymer gels, a possible involvement of swelling of the gel-like cytoskeleton structures in electrical regulation in the cell was postulated.

Quantal Sarcomere-Length Changes in Relaxed Single Myofibrils

We carried out experiments on single isolated myofibrils in which thin filaments had been functionally removed, leaving the connecting (titin) filaments as the sole agent taking up the length change. With technical advances that gave sub-nanometer detectability we examined the time course of single sarcomere-length change when the myofibril was ramp-released or ramp-stretched by a motor. The sarcomere-length change was stepwise. Step sizes followed a consistent pattern: the smallest was approximately 2.3 nm, and others were integer multiples of that value. The approximately 2.3-nm step quantum is the smallest consistent biomechanical event ever demonstrated. Although the length change must involve the connecting filament, the size of the quantum is an order of magnitude smaller than anticipated from folding of Ig- or fibronectin-like domains, implying either that folding occurs in sub-domain units or that other mechanisms are involved.

Quantal length changes in single contracting sarcomeres

The time course of shortening was investigated in the single sarcomere, the smallest contractile unit that retains natural structure. We projected the striation patterns of single bumblebee flight-muscle myofibrils onto a linear photodiode array, which was scanned periodically to produce repetitive traces of intensity vs. position along the array. Sarcomere length was taken as the span between adjacent A-band or Z-line centroids. When myofibrils were ramp-released by a motor, individual sarcomeres shortened in steps punctuated by pauses. The single sarcomere-shortening trace was consistently stepwise both in activated and relaxed specimens. Although step size was variable, the size distribution showed a signature-like feature: the histogram comprised distinct peaks that were spaced quasi-regularly. In the activated myofibrils the interpeak separation corresponded to 2.71 nm per half-sarcomere. This value is equal to the linear advance of actin subunits along the thin filament. Thus, actin filaments translate over thick filaments by steps that may be integer multiples of the actin-subunit spacing.

Mechanoelectric potentials in synthetic hydrogels: Possible relation to cytoskeleton

Mechanical and electrical properties of a synthetic polyelectrolyte hydrogel considered as a model of the cytoskeletal gel were studied. Hydrogels were synthesized from polymethacrylic acid by radical polymerization in aqueous solution. The electrical charge was introduced into the gel network by partial neutralization of monomer acids with magnesium hydroxide. Through the use of a motor, triangular longitudinal (axial) deformations were applied to gel samples. Simultaneously, the electrochemical (Donnan) potential of the gel was measured using conventional microelectrodes. We found that: (1) the Young modulus of the gel is 0.53 kPa; (2) at a given deformation velocity, the extent of gel deformation closely correlates with the gel potential; and (3) at the same level of gel deformation, the lower the deformation velocity, the higher the relative change of gel potential. These findings show a striking similarity to the data obtained in living cells, particularly in cardiac myocytes. A hypothesis involving the deformation-induced solvent migration from the gel to the surrounding solution is considered. It is concluded that the physicochemical features of the cytoskeletal gel may play a role in determining the mechanoelectric properties of excited cells.

Effect of sarcomere length on step size in relaxed rabbit psoas muscle

Recent experiments have shown that shortening and stretching of sarcomeres in single activated and unactivated myofibrils occur in stepwise fashion (Yang et al.(1998)Biophys J74: 1473–1483; Blyakhman et al.(2001)Biophys J81: 1093–1100; Yakovenko et al.(2002)Am J Physiol Cell Physiol283: 735–742). Here, we carried out measurements on single myofibrils from rabbit psoas muscle to investigate steps in unactivated specimens in more detail. Activated and unactivated myofibrils were released and stretched in ramp-like fashion. The time course of length change in the single sarcomere was consistently stepwise. We found that in the unactivated myofibrils, step size depended on initial sarcomere length, diminishing progressively with increase of initial sarcomere length, whereas in the case of activated sarcomeres, step size was consistently 2.7 nm.

Why the left ventricle is not a sphere

In this study, we have tried to understand why the left ventricle LV is not a homogeneous sphere. An experimental model of a spherical ventricle was developed. The chamber was configured as a mathematical model, and the wall properties were represented by isolated cardiac muscles. The stroke work of the spherical LV when modelling different types of inhomogeneity in the wall structure was investigated. It was found that the emergence of even slight inhomogeneity in a spherical ventricle inevitably results in a diminution of pump function. It was concluded that at a given level of the myocardial contractility, a homogeneous spherical LV would not have any functional reserve, ie no ability to maintain pump function in case of additional load. Functional reserve can be achieved only with a certain degree of inhomogeneity. Thus, inhomogeneity in the normal left ventricular wall structure constitutes a strategic functional reserve that is absent in a homogeneous spherical ventricle.

Polyacrylamide Ferrogels with Magnetite or Strontium Hexaferrite: Next Step in the Development of Soft Biomimetic Matter for Biosensor Applications

Magnetic biosensors are an important part of biomedical applications of magnetic materials. As the living tissue is basically a “soft matter.” this study addresses the development of ferrogels (FG) with micron sized magnetic particles of magnetite and strontium hexaferrite mimicking the living tissue. The basic composition of the FG comprised the polymeric network of polyacrylamide, synthesized by free radical polymerization of monomeric acrylamide (AAm) in water solution at three levels of concentration (1.1 M, 0.85 M and 0.58 M) to provide the FG with varying elasticity. To improve FG biocompatibility and to prevent the precipitation of the particles, polysaccharide thickeners—guar gum or xanthan gum were used. The content of magnetic particles in FG varied up to 5.2 wt % depending on the FG composition. The mechanical properties of FG and their deformation in a uniform magnetic field were comparatively analyzed. FG filled with strontium hexaferrite particles have larger Young’s modulus value than FG filled with magnetite particles, most likely due to the specific features of the adhesion of the network’s polymeric subchains on the surface of the particles. FG networks with xanthan are stronger and have higher modulus than the FG with guar. FG based on magnetite, contract in a magnetic field 0.42 T, whereas some FG based on strontium hexaferrite swell. Weak FG with the lowest concentration of AAm shows a much stronger response to a field, as the concentration of AAm governs the Young’s modulus of ferrogel. A small magnetic field magnetoimpedance sensor prototype with Co68.6Fe3.9Mo3.0Si12.0B12.5 rapidly quenched amorphous ribbon based element was designed aiming to develop a sensor working with a disposable stripe sensitive element. The proposed protocol allowed measurements of the concentration dependence of magnetic particles in gels using magnetoimpedance responses in the presence of magnetite and strontium hexaferrite ferrogels with xanthan. We have discussed the importance of magnetic history for the detection process and demonstrated the importance of remnant magnetization in the case of the gels with large magnetic particles.

Mechanical, Electrical and Magnetic Properties of Ferrogels with Embedded Iron Oxide Nanoparticles Obtained by Laser Target Evaporation: Focus on Multifunctional Biosensor Applications

Hydrogels are biomimetic materials widely used in the area of biomedical engineering and biosensing. Ferrogels (FG) are magnetic composites capable of functioning as magnetic field sensitive transformers and field assisted drug deliverers. FG can be prepared by incorporating magnetic nanoparticles (MNPs) into chemically crosslinked hydrogels. The properties of biomimetic ferrogels for multifunctional biosensor applications can be set up by synthesis. The properties of these biomimetic ferrogels can be thoroughly controlled in a physical experiment environment which is much less demanding than biotests. Two series of ferrogels (soft and dense) based on polyacrylamide (PAAm) with different chemical network densities were synthesized by free-radical polymerization in aqueous solution with N,N’-methylene-diacrylamide as a cross-linker and maghemite Fe2O3 MNPs fabricated by laser target evaporation as a filler. Their mechanical, electrical and magnetic properties were comparatively analyzed. We developed a giant magnetoimpedance (MI) sensor prototype with multilayered FeNi-based sensitive elements deposited onto glass or polymer substrates adapted for FG studies. The MI measurements in the initial state and in the presence of FG with different concentrations of MNPs at a frequency range of 1–300 MHz allowed a precise characterization of the stray fields of the MNPs present in the FG. We proposed an electrodynamic model to describe the MI in multilayered film with a FG layer based on the solution of linearized Maxwell equations for the electromagnetic fields coupled with the Landau-Lifshitz equation for the magnetization dynamics.

Mechanical characteristics of synthetic polyelectrolyte gel as a physical model of the cytoskeleton

A physical model of the cytoskeleton based on synthetic polyelectrolyte hydrogel of polymethacrylic acid has been proposed. From the physicochemical point of view, the structures of polyelectrolyte gel and the cytoskeleton show a high degree of similarity. It has been shown that polyelectrolyte gel can shorten and produce mechanical stress in response to changes in the composition of the surrounding solution. The mechanical properties of the model gel have been evaluated: Young modulus (2–6 kPa), stress relaxation time (0.1–1 s), and apparent viscosity (0.3–3 kPa s). The viscoelastic properties of the gel depend on the degree of its swelling. It has been demonstrated that the mechanical properties of gels of polymethacrylic acid are close to those of biological objects.

Sarcomere Dynamics, Stepwise Shortening and the Nature of Contraction

This paper has a dual goal. First it outhnes the methods that have evolved to track the time course of sarcomere length (SL) with increasingly high precision. Serious attempts at this began roughly at the time the sliding filament theory was introduced in the mid-1950s, and have progressed to the point where resolution has reached the nanometer level. Second, and within the context of these developments, it considers one of the more controversial aspects of these developments: stepwise shortening. Stepwise shortening was first observed a quarter century ago (Pollack et al., 1977), and has provided no shortage of controversy for the muscle-contraction field.