Simultaneous Application of Interstitial Flow and Cyclic Mechanical Strain to a Three-Dimensional Cell-Seeded Hydrogel

The present study describes the design and validation of a simple apparatus to apply simultaneous mechanical and fluidic stress to three-dimensional (3D) cell-seeded collagen hydrogels. Constructs were formed in wells in a silicone substrate that could be stretched cyclically, and were also fitted with inlet ports to apply fluid flow. Acid etching was used to retain adhesion of the gels to the walls of the well, and an acellular layer of collagen hydrogel was used to distribute flow evenly. Finite element modeling showed that 5% uniaxial strain applied to the entire silicone substrate resulted in similar to 6.5% strain in each of the gel constructs. Permeability testing and flow observation showed that acellular hydrogels were fourfold more permeable than cardiac fibroblast-seeded gels, and that the fluid distributed evenly in the acellular layer before entering the cell-seeded gel. Viability testing and imaging demonstrated that cells remained viable with expected fibroblast morphology for the 120 h duration of the experiments. These results demonstrate that this simple bioreactor can be used to study the effects of mechanical strain and interstitial flow in 3D protein hydrogels. Such 3D tissue models have utility in studying cell and tissue responses to their mechanical environment.