Biomedical Microdevices

, Volume 12, Issue 6, pp 1073–1085 | Cite as

Hypertrophy, gene expression, and beating of neonatal cardiac myocytes are affected by microdomain heterogeneity in 3D

  • Matthew W. Curtis
  • Sadhana Sharma
  • Tejal A. Desai
  • Brenda Russell
Article

Abstract

Cardiac myocytes are known to be influenced by the rigidity and topography of their physical microenvironment. It was hypothesized that 3D heterogeneity introduced by purely physical microdomains regulates cardiac myocyte size and contraction. This was tested in vitro using polymeric microstructures (G′ = 1.66 GPa) suspended with random orientation in 3D by a soft Matrigel matrix (G′ = 22.9 Pa). After 10 days of culture, the presence of 100 μm-long microstructures in 3D gels induced fold increases in neonatal rat ventricular myocyte size (1.61 ± 0.06, p < 0.01) and total protein/cell ratios (1.43 ± 0.08, p < 0.05) that were comparable to those induced chemically by 50 μM phenylephrine treatment. Upon attachment to microstructures, individual myocytes also had larger cross-sectional areas (1.57 ± 0.05, p < 0.01) and higher average rates of spontaneous contraction (2.01 ± 0.08, p < 0.01) than unattached myocytes. Furthermore, the inclusion of microstructures in myocyte-seeded gels caused significant increases in the expression of beta-1 adrenergic receptor (β1-AR, 1.19 ± 0.01), cardiac ankyrin repeat protein (CARP, 1.26 ± 0.02), and sarcoplasmic reticulum calcium-ATPase (SERCA2, 1.59 ± 0.12, p < 0.05), genes implicated in hypertrophy and contractile activity. Together, the results demonstrate that cardiac myocyte behavior can be controlled through local 3D microdomains alone. This approach of defining physical cues as independent features may help to advance the elemental design considerations for scaffolds in cardiac tissue engineering and therapeutic microdevices.

Keywords

Cardiomyocyte Beat frequency Cell remodeling Focal adhesion Mechanotransduction Microstructure Microenvironment Three dimensions Hypertrophy Spontaneous contraction 

Abbreviations

3D

three dimensions

AraC

cytosine β-D-arabino-furanoside

β1-AR

beta-1 adrenergic receptor

β2M

beta-2 microglobulin

BDM

2,3-butanedione monoxime

BPM

beats per minute

BSA

bovine serum albumin

CARP

cardiac ankyrin repeat protein

COX8H

cytochrome c oxidase subunit VIII heart/muscle

DAPI

4′,6-diamidino-2-phenylindole

DIC

differential interference contrast

DMEM

Dulbecco’s modified Eagle’s medium

DOB

dobutamine

ECM

extracellular matrix

PBS

phosphate buffered saline

PE

phenylephrine

PEGDMA

poly(ethylene glycol) dimethacrylate

SDS

sodium dodecyl sulfate

SEM

standard error of measurement

SERCA2

sarcoplasmic reticulum calcium-ATPase

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Matthew W. Curtis
    • 1
  • Sadhana Sharma
    • 2
  • Tejal A. Desai
    • 3
  • Brenda Russell
    • 2
  1. 1.Department of BioengineeringUniversity of Illinois at ChicagoChicagoUSA
  2. 2.Department of Physiology and Biophysics (MC 901)University of Illinois at ChicagoChicagoUSA
  3. 3.Department of Physiology and Division of BioengineeringUniversity of California at San FranciscoSan FranciscoUSA

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