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Biomechanics and Modeling in Mechanobiology

, Volume 12, Issue 1, pp 95–109 | Cite as

Microdomain heterogeneity in 3D affects the mechanics of neonatal cardiac myocyte contraction

  • Matthew W. Curtis
  • Elisa Budyn
  • Tejal A. Desai
  • Allen M. Samarel
  • Brenda RussellEmail author
Original Paper

Abstract

Cardiac muscle cells are known to adapt to their physical surroundings, optimizing intracellular organization and contractile function for a given culture environment. A previously developed in vitro model system has shown that the inclusion of discrete microscale domains (or microrods) in three dimensions (3D) can alter long-term growth responses of neonatal ventricular myocytes. The aim of this work was to understand how cellular contact with such a domain affects various mechanical changes involved in cardiac muscle cell remodeling. Myocytes were maintained in 3D gels over 5 days in the presence or absence of 100−μm-long microrods, and the effect of this local heterogeneity on cell behavior was analyzed via several imaging techniques. Microrod abutment resulted in approximately twofold increases in the maximum displacement of spontaneously beating myocytes, as based on confocal microscopy scans of the gel xy-plane or the myocyte long axis. In addition, microrods caused significant increases in the proportion of aligned myofibrils (≤20° deviation from long axis) in fixed myocytes. Microrod-related differences in axial contraction could be abrogated by long-term interruption of certain signals of the RhoA-/Rho-associated kinase (ROCK) or protein kinase C (PKC) pathway. Furthermore, microrod-induced increases in myocyte size and protein content were prevented by ROCK inhibition. In all, the data suggest that microdomain heterogeneity in 3D appears to promote the development of axially aligned contractile machinery in muscle cells, an observation that may have relevance to a number of cardiac tissue engineering interventions.

Keywords

Mechanobiology Muscle hypertrophy Digital image correlation Microenvironment Finite element Mechanotransduction 

Abbreviations

3D

Three dimensions

AraC

Cytosine β-D-arabino-furanoside

BSA

Bovine serum albumin

DAPI

4′,6-Diamidino-2-phenylindole

DMEM

Dulbecco’s modified Eagle’s medium

DMSO

Dimethyl sulfoxide

ECL

Enhanced chemiluminescence

ECM

Extracellular matrix

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

HRP

Horseradish peroxidase

MLC

Myosin light chain

MLCK

Myosin light chain kinase

MLCP

Myosin light chain phosphatase

PBS

Phosphate-buffered saline

PKC

Protein kinase C

PVDF

Polyvinylidene fluoride

RACK1

Receptor for activated C-kinase-1

ROCK

Rho-associated kinase

SEM

Standard error of measurement

TBST

Tris-buffered saline-Tween 20

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Matthew W. Curtis
    • 1
  • Elisa Budyn
    • 2
  • Tejal A. Desai
    • 3
  • Allen M. Samarel
    • 4
  • Brenda Russell
    • 1
    Email author
  1. 1.Department of Physiology and BiophysicsUniversity of Illinois at ChicagoChicagoUSA
  2. 2.Department of Mechanical and Industrial EngineeringUniversity of Illinois at ChicagoChicagoUSA
  3. 3.Department of Physiology and Division of BioengineeringUniversity of California at San FranciscoSan FranciscoUSA
  4. 4.The Cardiovascular InstituteLoyola University Medical CenterMaywoodUSA

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