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A Fluorescent Intravital Imaging Approach to Study Load-Induced Calcium Signaling Dynamics in Mouse Osteocytes
Published on: February 24, 2023
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3D osteocyte networks under Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS)
Biorxiv : the Preprint Server for Biology
|July 14, 2025
Summary
This study introduces a novel microfluidic model to investigate how pulsatile mechanical stimuli affect 3D osteocyte networks. The model demonstrates that Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS) can trigger mechanotransduction and maintain osteocyte gene expression.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Skeletal Mechanobiology
Background:
- Osteocytes are crucial for skeletal adaptation to mechanical forces.
- Existing in vitro models are limited in simulating dynamic mechanical stimuli on 3D osteocyte networks.
Purpose of the Study:
- To develop and validate a microfluidic model for studying 3D osteocyte networks under pulsatile mechanical stimulation.
- To investigate the effects of Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS) on osteocyte behavior and signaling.
Main Methods:
- Fabrication of a multi-chambered PDMS microfluidic chip using digital light projection stereolithography.
- Encapsulation of murine MLO-Y4 osteocytes in a collagen matrix to form 3D networks.
- Application of daily PUFFS for up to 21 days, combined with experimental, computational, and analytical characterization.
Main Results:
- PUFFS at 0.33 and 1.66 Hz induced mechanotransduction through calcium signaling propagated via Cx43 junctions.
- Osteocytes cultured in the model maintained expression of key osteocyte genes for 21 days.
- The model successfully characterized mechanical environments and cellular responses.
Conclusions:
- The developed microfluidic model provides a robust platform for studying 3D osteocyte networks under dynamic mechanical loading.
- PUFFS can effectively stimulate mechanotransduction pathways in osteocytes.
- This model serves as a valuable testbed for understanding skeletal tissue responses to mechanical stimuli.
