A novel micro tensile tester with feed-back control for viscoelastic analysis of single isolated smooth muscle cells.

A novel micro tensile tester was developed to measure the viscoelastic properties of single isolated cells. A specimen cell was held with two glass micropipettes, the tips of which were coated with a urethane resin adhesive. One pipette was rigid and fixed in position, and the other was deflectable and could be moved by a laboratory-made piezoelectric actuator to stretch the specimen. The actuator was made of an originally designed lever mechanism which can magnify the deformation of conventional piezoceramic actuator and allowed for sufficient displacement up to 150 microm. The distance between the two pipette tips was obtained by real-time image processing and kept constant following stepwise stretching with an accuracy of +/-0.1 microm by changing the voltage applied to the piezoceramics under feed-back control. The force applied to the cell was measured by the deflection of a cantilever part of the deflectable pipette. The stress relaxation test was performed for cultured rat aortic smooth muscle cells. The cells were stretched by 75% and their length was kept constant. Viscoelastic analysis with a standard linear solid showed that the relaxation time constant of the isolated smooth muscle cells was 250+/-42 s (n=8, mean+/-S.E.M.) which was over six times longer than that reported for vascular endothelial cells and fibroblasts.

[1]  R M Nerem,et al.  Application of the micropipette technique to the measurement of cultured porcine aortic endothelial cell viscoelastic properties. , 1990, Journal of biomechanical engineering.

[2]  K. Jacobson,et al.  Local measurements of viscoelastic parameters of adherent cell surfaces by magnetic bead microrheometry. , 1998, Biophysical journal.

[3]  S Sugiura,et al.  A novel method to study contraction characteristics of a single cardiac myocyte using carbon fibers. , 2001, American journal of physiology. Heart and circulatory physiology.

[4]  H. Miyazaki,et al.  A newly designed tensile tester for cells and its application to fibroblasts. , 2000, Journal of biomechanics.

[5]  M. Mulvany,et al.  Mechanical and Morphological Properties of Arterial Resistance Vessels in Young and Old Spontaneously Hypertensive Rats , 1979, Circulation research.

[6]  Julie H. Campbell,et al.  Vascular smooth muscle in culture , 1987 .

[7]  T Matsumoto,et al.  Mechanical and dimensional adaptation of rat aorta to hypertension. , 1994, Journal of biomechanical engineering.

[8]  Takeo Matsumoto,et al.  Effect of actin filament distribution on tensile properties of smooth muscle cells obtained from rat thoracic aortas. , 2006, Journal of biomechanics.

[9]  T Matsumoto,et al.  Stress and strain distribution in hypertensive and normotensive rat aorta considering residual strain. , 1996, Journal of biomechanical engineering.

[10]  G. Shue,et al.  The frequency response of smooth muscle stiffness during Ca2+-activated contraction. , 1999, Biophysical journal.

[11]  F Guilak,et al.  Viscoelastic properties of chondrocytes from normal and osteoarthritic human cartilage , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  N Wang,et al.  Mechanical interactions among cytoskeletal filaments. , 1998, Hypertension.

[13]  H. Wolinsky,et al.  Long‐Term Effects of Hypertension on the Rat Aortic Wall and Their Relation to Concurrent Aging Changes: MORPHOLOGICAL AND CHEMICAL STUDIES , 1972, Circulation research.

[14]  Joachim P Spatz,et al.  Mechanical response analysis and power generation by single-cell stretching. , 2005, Chemphyschem : a European journal of chemical physics and physical chemistry.

[15]  N. Caille,et al.  Contribution of the nucleus to the mechanical properties of endothelial cells. , 2002, Journal of biomechanics.

[16]  Masaaki Sato,et al.  Smooth muscle cells freshly isolated from rat thoracic aortas are much stiffer than cultured bovine cells : Possible effect of phenotype , 2000 .

[17]  R. Skalak,et al.  Passive mechanical properties of human leukocytes. , 1981, Biophysical journal.

[18]  P. Smith,et al.  Selected contribution: mechanical strain increases force production and calcium sensitivity in cultured airway smooth muscle cells. , 2000, Journal of applied physiology.

[19]  W. Bayliss.  On the local reactions of the arterial wall to changes of internal pressure , 1902, The Journal of physiology.

[20]  R. Cox.  Comparison of arterial wall mechanics in normotensive and spontaneously hypertensive rats. , 1979, American Journal of Physiology.

[21]  Takeo Matsumoto,et al.  DEVELOPMENT OF A NOVEL MICRO TENSILE TESTER FOR SINGLE ISOLATED CELLS AND ITS APPLICATION TO VISCOELASTIC ANALYSIS OF AORTIC SMOOTH MUSCLE CELLS , 2005 .

[22]  R. Hochmuth,et al.  Red cell extensional recovery and the determination of membrane viscosity. , 1979, Biophysical journal.

[23]  Takeo Matsumoto,et al.  Mechanical Anisotropy of Rat Aortic Smooth Muscle Cells Decreases with Their Contraction , 2004 .

[24]  O. Thoumine,et al.  Time scale dependent viscoelastic and contractile regimes in fibroblasts probed by microplate manipulation. , 1997, Journal of cell science.