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Probing cellular response to topography in three dimensions.

Colin D Paul1, Alex Hruska1, Jack R Staunton1

  • 1Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, USA.

Biomaterials
|January 15, 2019
PubMed
Summary

Engineered 3D matrices mimic tissue microenvironments, revealing that myosin II regulates cell alignment to topographical cues in engineered matrices and in vivo. This finding advances understanding of cell-matrix interactions.

Keywords:
Cell alignmentCell protrusionsEngineered matricesMicrorheologyPhysical propertiesTopographical cues

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Area of Science:

  • Biophysics
  • Biomaterials Science
  • Cell Biology

Background:

  • Tissue microenvironments feature mechanical properties, fibrillar alignment, and extracellular matrix (ECM) architecture.
  • These biophysical cues, along with chemical signals, drive cellular responses.

Purpose of the Study:

  • To engineer 3D fibrillar architectures using magnetic assembly of functionalized colloidal particles.
  • To investigate cell behavior in response to defined fibrillar geometries and ECM proteins.
  • To examine factors influencing cell alignment to topographical cues in 3D matrices.

Main Methods:

  • Bottom-up assembly of paramagnetic colloidal particles functionalized with ECM proteins.
  • Creation of 3D amorphous hydrogels with controlled fibrillar architecture.
  • Culturing multiple human cell lines and assessing cell protrusion and alignment.
  • Measuring focal adhesion protein localization and assessing the role of specific proteins (integrin β1, fascin, myosin II).
  • In vivo validation using blebbistatin treatment.

Main Results:

  • Resolved spatial heterogeneities in microscale mechanical properties near aligned fibers.
  • Cells extended protrusions parallel or perpendicular to aligned fibers, irrespective of ECM coating.
  • Integrin β1 and fascin influenced protrusion extension but not alignment.
  • Myosin II inhibition disrupted cell alignment to topographical cues in 3D matrices and in vivo.

Conclusions:

  • Myosin II plays a critical role in sensing topographical cues within 3D engineered matrices.
  • This myosin II dependence is conserved in vivo, affecting cell alignment along blood vessels.
  • The engineered platform allows for detailed study of cell-matrix interactions and mechanosensing.