Design and characterization of a modified T-flask bioreactor for continuous monitoring of engineered tissue stiffness.
Controlling environmental conditions, such as mechanical stimuli, is critical for directing cells into functional tissue. This study reports on the development of a bioreactor capable of controlling the mechanical environment and continuously measuring force-displacement in engineered tissue. The bioreactor was built from off the shelf components, modified off the shelf components, and easily reproducible custom built parts to facilitate ease of setup, reproducibility and experimental flexibility. A T-flask was modified to allow for four tissue samples, mechanical actuation via a LabView controlled stepper motor and transduction of force from inside the T-flask to an external sensor. In vitro bench top testing with instrumentation springs and tissue culture experiments were performed to validate system performance. Force sensors were highly linear (R(2) > 0.998) and able to maintain force readings for extended periods of time. Tissue culture experiments involved cyclic loading of polyurethane scaffolds seeded with and without (control) human foreskin fibroblasts for 8 h/day for 14 days. After supplementation with TGF-beta, tissue constructs showed an increase in stiffness between consecutive days and from the acellular controls. These experiments confirmed the ability of the bioreactor to distinguish experimental groups and monitor tissue stiffness during tissue development.
Lasher
Richard A
RA
Dept. of Bioengineering, University of Utah, Salt Lake City, UT 84112, USA.
Wolchok
Jeffrey C
JC
Parikh
Monir K
MK
Kennedy
James P
JP
Hitchcock
Robert W
RW
D019149:Bioreactors; D004548:Elasticity; D004867:Equipment Design; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D015203:Reproducibility of Results; D046509:Tissue Culture Techniques / Q000295:instrumentation* / Q000379:methods; D023822:Tissue Engineering; D054457:Tissue Scaffolds