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

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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.

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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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Authors and Affiliations

  1. Department of Physiology and Biophysics, University of Illinois at Chicago, 835 South Wolcott Avenue, Chicago, IL, 60612, USA

    Matthew W. Curtis & Brenda Russell

  2. Department of Mechanical and Industrial Engineering, University of Illinois at Chicago, Chicago, IL, USA

    Elisa Budyn

  3. Department of Physiology and Division of Bioengineering, University of California at San Francisco, San Francisco, CA, USA

    Tejal A. Desai

  4. The Cardiovascular Institute, Loyola University Medical Center, Maywood, IL, USA

    Allen M. Samarel

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  1. Matthew W. Curtis
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  2. Elisa Budyn
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  3. Tejal A. Desai
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  4. Allen M. Samarel
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  5. Brenda Russell
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Correspondence to Brenda Russell.

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Curtis, M.W., Budyn, E., Desai, T.A. et al. Microdomain heterogeneity in 3D affects the mechanics of neonatal cardiac myocyte contraction. Biomech Model Mechanobiol 12, 95–109 (2013). https://doi.org/10.1007/s10237-012-0384-9

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  • DOI: https://doi.org/10.1007/s10237-012-0384-9

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