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3D Osteocyte Networks under Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS).

Anna-Blessing Merife1, Arun Poudel1, Angelika Polshikova1

  • 1Department of Chemical and Biomedical Engineering, L.C. Smith College of Engineering Syracuse University, Syracuse, New York 13244, United States.

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Summary

This study introduces a novel microfluidic model to investigate how pulsatile mechanical stimuli affect three-dimensional (3D) osteocyte networks, revealing insights into skeletal mechanotransduction and cell communication.

Keywords:
3D culturesMLO-Y4chipin vitro modelmechanical stimulimicrofluidicosteocytes

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Skeletal Mechanobiology

Background:

  • Osteocytes are crucial for skeletal mechanoadaptation.
  • Limited in vitro models exist for studying 3D osteocyte networks under dynamic mechanical forces.

Purpose of the Study:

  • To develop and validate a microfluidic in vitro model for 3D osteocyte networks subjected to Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS).
  • To investigate the cellular and molecular responses of osteocytes to dynamic mechanical stimulation.

Main Methods:

  • Fabrication of a polydimethylsiloxane (PDMS) microfluidic chip using digital light projection stereolithography.
  • Encapsulation of murine MLO-Y4 osteocytes within a collagen matrix to form 3D networks.
  • Application of daily PUFFS for up to 21 days, coupled with experimental, computational, and analytical characterization.

Main Results:

  • PUFFS at 0.33 and 1.66 Hz induced mechanotransduction via calcium signaling propagated through Cx43 junctions.
  • Osteocytes maintained expression of key genes for up to 21 days under PUFFS.
  • The model successfully characterized the mechanical environment and cellular responses.

Conclusions:

  • The developed microfluidic model provides a platform for studying 3D osteocyte network responses to dynamic mechanical stimuli.
  • This model is valuable for understanding mechanotransduction in skeletal tissues.