Key Findings
The combined application of fluid shear stress and intermittent hydrostatic pressure enhances the osteogenic differentiation of bone marrow stromal cells more effectively than direct perfusion alone. In a seven-day culture using the Y201 clonal line of human telomerase reverse transcriptase-immortalized bone marrow-derived mesenchymal stromal cells, the dual-stimulus environment promoted osteogenic lineage progression without compromising cell viability. Resazurin reduction assays indicated that both perfusion conditions significantly increased metabolic activity compared to static cultures.
Technical Details
The device consists of three independent culture chambers in closed loops, powered by a multichannel peristaltic pump with flow rates from 0.024 to 24 milliliters per minute. To generate intermittent hydrostatic pressure, a three-way solenoid pinch valve periodically blocks the outflow; when closed, pressure climbs within the chamber until the valve releases. An Arduino-based controller allows users to set stimulation periods from 0.25 to 30 seconds and duty cycles from 1 to 100 percent.
The system was characterized using two 3D-printed polylactic acid (PLA) scaffolds:
- PLA600: Mimics the human iliac crest (60% porosity, 350 micrometer trabeculae, 531 micrometer spacing), generating peak pressures from 1.81 to 15.09 kilopascals.
- P3S3: Mimics jawbone architecture (44.7% porosity, 258 micrometer trabeculae, 284 micrometer spacing), generating peak pressures from 4.53 to 30.59 kilopascals.
Calculated shear stress ranged from 0.48 to 481.99 millipascals for PLA600 and 0.68 to 683.26 millipascals for P3S3.
Background and Significance
Bone tissue is naturally subjected to both hydrostatic pressure and fluid flow during physical activity, with in vivo pressures typically ranging from 3 to 8 kilopascals during daily activity and up to 35 kilopascals during intense exercise. Previous laboratory devices often applied only one stimulus or were limited to 2D cell layers. By recreating these dual mechanical loads in a 3D environment, this bioreactor provides a more biomimetic platform for studying bone tissue development and regeneration.
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