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Tunable photoinitiated hydrogel microspheres for quantifying cell-generated forces in complex three-dimensional

Antoni Garcia-Herreros1, Yi-Ting Yeh2, Yunpeng Tu3

  • 1Department of Mechanical and Aerospace Engineering, University of California, San Diego, La Jolla, CA, United States.

Acta Biomaterialia
|August 29, 2025
PubMed
Summary

We developed a microfluidic method to create tunable, uniform cellular force microscopy probes. This technique quantifies cellular forces in complex environments, revealing endothelial cell compression during engulfment.

Keywords:
Cell mechanicsCellular force probesMechanobiologyMicrofabricationMicrofluidicsPolyacrylamideThree dimensional hydrogelsTraction force microscopy

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Area of Science:

  • Cell Biology
  • Biophysics
  • Materials Science

Background:

  • Quantifying mechanical forces in cell biology is vital but challenging in native environments.
  • Hydrogel microspheres are promising for force measurement but face production limitations.
  • Existing methods struggle with high-yield, tunable, and monodisperse probe fabrication.

Purpose of the Study:

  • To develop a high-throughput microfluidic method for producing cellular force microscopy probes.
  • To create probes with controlled size, elastic modulus, and tunable properties.
  • To enable accurate measurement of cellular forces in physiologically relevant systems.

Main Methods:

  • Utilized flow-focusing microfluidics for reproducible droplet generation.
  • Employed acrylamide precursor and LAP photoinitiator for controlled polymerization.
  • Functionalized microspheres with ECM proteins and embedded fluorescent nanobeads for tracking.

Main Results:

  • Generated large quantities of monodisperse polyacrylamide (PAAm) hydrogel microspheres.
  • Successfully functionalized probes and embedded tracking nanobeads.
  • Measured 3D traction forces exerted by vascular endothelial cells on probes.

Conclusions:

  • The microfluidic method overcomes barriers in PAAm microsphere fabrication.
  • The developed probes offer tunable stiffness and monodisperse size.
  • This technique provides a powerful tool for studying cellular mechanobiology and forces in complex systems.