Biomedical subjects

R E Waugh

Publications and source records attributed to R E Waugh.

43 records · Page 3Linked to original sources

PubMed1983

Effects of abnormal cytoskeletal structure on erythrocyte membrane mechanical properties.

Measurements of the mechanical properties of the erythrocyte membrane provide a direct assessment of the proper function of its structural components. To assess the effects of alterations in molecular structure on membrane mechanical properties, measurements have been performed on cells from six individuals whose membranes contain inherited, biochemically characterized structural defects. Because the contribution of the membrane skeleton to the mechanical behavior of the membrane is most evident in shear deformation, mechanical experiments were performed to measure the material constants which characterize the response of the membrane to shear force resultants. The surface elastic shear modulus characterizes the elastic response of the membrane; the yield shear resultant is the maximum shear force resultant which the membrane can support elastically; and the plastic viscosity coefficient characterizes the rate of membrane deformation when the elastic limit has been exceeded. Generally, it was found that when the molecular defect is found to occur in a region of the skeleton which is stress-supporting, the maximum elastic strength of the membrane is reduced. However, the magnitude of the reduction can be quite different for membranes having similar or even identical defects. In some cases the differences can be attributed to the removal of the most fragile cells of the population by the spleen, but other results indicate that the biochemical description of the defects may be incomplete. These results emphasize the need for further refinements both in the biochemical characterization of membrane skeleton structure and in the description and measurement of membrane mechanical properties.

Biomechanical Phenomena↗

PubMed1982

Temperature dependence of the yield shear resultant and the plastic viscosity coefficient of erythrocyte membrane. Implications about molecular events during membrane failure.

Structural failure of the erythrocyte membrane in shear deformation occurs when the maximum shear resultant (force/length) exceeds a critical value, the yield shear resultant. When the yield shear resultant is exceeded, the membrane flows with a rate of deformation characterized by the plastic viscosity coefficient. The temperature dependence of the yield shear resultant and the plastic viscosity coefficient have been measured over the temperature range 10-40 degrees C. Over this range the yield shear resultant does not change significantly (+/- 15%), but the plastic viscosity coefficient changes exponentially from a value of 1.3 X 10(-2) surface poise (dyn s/cm) at 10 degrees C to a value of 6.2 X 10(-4) surface poise (SP) at 40 degrees C. The different temperature dependence of these two parameters is not surprising, inasmuch as they characterize different molecular events. The yield shear resultant depends on the number and strength of intermolecular connections within the membrane skeleton, whereas the plastic viscosity depends on the frictional interactions between molecular segments as they move past one another in the flowing surface. From the temperature dependence of the plastic viscosity, a temperature-viscosity coefficient, E, can be calculated: eta p = constant X exp(--E/RT). This quantity (E) is related to the probability that a molecular segment can "jump" to its next location in the flowing network. The temperature-viscosity coefficient for erythrocyte membrane above the elastic limit is calculated to be 18 kcal/mol, which is similar to coefficients for other polymeric materials.

Erythrocyte Membrane↗

PubMed1982

Surface viscosity measurements from large bilayer vesicle tether formation. I. Analysis.

Recent observations indicate that it is possible to form tethers from large phospholipid vesicles. The process of tether formation is analyzed using a continuum mechanical approach to obtain the surface viscosity of the bilayer in terms of experimentally measurable parameters. The membrane is treated as a two-dimensional isotropic material which deforms a constant area. The constitutive equation relates the maximum surface shear resultant to the rate of deformation via the surface viscosity coefficient. The force which acts to increase the tether length is generated by fluid moving past the vesicle. The magnitude of the force is estimated from Stoke's drag equation. The analysis predicts that there is a critical force necessary to produce an increase in the tether length. A dimensionless tether growth parameter is defined, and its value is obtained as a function of the ratio of the applied force on the vesicle to the critical force. This relationship is independent of both the size of the vesicle and the radius of the tether. Knowing the force on the vesicle, the critical force, and the rate of tether growth, the surface viscosity can be calculated.

Lipid Bilayers↗

PubMed1982

Surface viscosity measurements from large bilayer vesicle tether formation. II. Experiments.

A mechanical experiment has been developed that measures an upper bound for the viscosity of a lipid bilayer membrane. In this experiment, strands of membrane (tethers) are formed from phospholipid vesicles attached to micropipettes by subjecting the vesicles to fluid drag. The rate of tether formation is measured as a function of the velocity of the suspending fluid. The surface viscosity can be calculated from this data using a theoretical relationship derived in a companion paper. Because of the multilamellar character of the vesicles, these values provide an upper bound for the viscosity of a single bilayer. The smallest values obtained in these measurements fell in the range 5.0-13.0 x 10(-6) dyn s/cm. These values are in relatively good agreement with the values calculated from lateral and rotational mobility measurements.

Lipid Bilayers↗

PubMedDate not supplied

Physical measurements of bilayer-skeletal separation forces.

Bilayer membranes are intrinsically fluid in character and require stabilization by association with an underlying cytoskeleton. Instability either in the membrane-associated cytoskeleton or in the association between the bilayer and the skeleton can lead to loss of membrane bilayer and premature cell death. In this report measurements of the physical strength of the association between membrane bilayer and the membrane-associated skeleton in red blood cells are reported. These measurements involve the mechanical formation of long, thin cylinders of membrane bilayer (tethers) from the red cell surface. Ultrastructural evidence is presented indicating that these tethers do not contain membrane skeleton and, furthermore, that they are deficient in at least some integral membrane proteins. By measuring the forces on the cell as the tether is formed and the dimensions of the tether, the energy associated with its formation can be calculated. The minimum force to form a tether was found to be approximately 50 pN corresponding to an energy of dissociation of 0.2-0.3 mJ/m2. Such measurements enable critical evaluation of potential physical mechanisms for the stabilization of the membrane bilayer by the underlying cytoskeleton. It is postulated that an important contribution to the energy of association between bilayer and skeleton comes from the increase in chemical potential due to the lateral segregation of lipids and integral proteins.

Biomechanical Phenomena↗

PubMedDate not supplied

A piconewton force transducer and its application to measurement of the bending stiffness of phospholipid membranes.

The bending stiffness of a phospholipid bilayer (Kc) was measured by forming thin bilayer cylinders (tethers) from giant phospholipid vesicles. Based on the balance of forces, the tether force was expected to be proportional to the square root of the membrane tension, with a constant of proportionality containing Kc. The membrane tension was controlled via the aspiration pressure in a micropipette used to hold the vesicle. The force on the tether was generated by an electromagnet acting on a paramagnetic bead attached to the vesicle surface. The magnitude of the force was determined from measurements of the magnet current, which was adjusted to maintain the position of the bead. Measurements were performed on vesicles composed of stearoyl-oleoyl-phosphatidylcholine plus 5% (by mole) biotinylated phosphatidylethanolamine to mediate adhesion to streptavidin-coated beads. From each vesicle, tethers were formed repeatedly at different values of the membrane tension. The expected relationship between membrane tension and tether force was observed. The mean value of Kc for 10 different vesicles was 1.17 x 10(-19) J (SD = 0.08 x 10(-19) J). The precision of these data demonstrates the reliability of this approach, which avoids uncertainties of interpretation and measurement that may be associated with other methods for determining Kc.

Calibration↗

PubMedDate not supplied

Changes in HL-60 cell deformability during differentiation induced by DMSO.

We have investigated changes in cellular deformability during promyelocytic leukemic HL-60 cell maturation. HL-60 cells were induced to mature with 1.25% dimethyl sulfoxide. Cellular deformability was evaluated by single-cell micropipette aspiration at one day, four days and seven days after induction. HL-60 cells were found to decrease in size and increase in deformability with maturation. When tested under the same aspiration pressures (0.5-1.3 kPa), cytoplasmic viscosity was found to vary from 210 to 85 Pa.s for cells prior to induction; it varied from 85 to 40 Pa.s for cells seven days after induction. Further, cytoplasmic viscosity exhibits power-law dependence on shear rate, mu = mu c (gamma m/gamma c)-b, where mu is cytoplasmic viscosity, gamma m is mean shear rate during cell entry, mu c is the characteristic viscosity at the characteristic shear rate, gamma c, and b is a material coefficient. Cells of all maturities showed similar dependence on shear rate (b approximately 0.5), but the characteristic viscosity decreased with maturation except for Day 1. When gamma c was set to 1 s-1, mu c = 236 +/- 5 Pa.s for cells prior to induction, mu c = 239 +/- 7, 209 +/- 7 and 175 +/- 14 Pa.s for cells on Days 1, 4 and 7 of induction, respectively.

Cell Differentiation↗