Stress stiffened silicon nitride micro bridges array as substrate with tunable stiffness for cell culture

Recently, interactions between one-dimensional structural stiffness of physical micro environments and cell biological process have been widely studied. However in previous studies, the influence of structural stiffness on biological process was coupled with the influence of micro fiber curvature. Therefore decoupling the influences of fiber curvature and structural stiffness on cell biological process is of prime importance. In this study, we proposed a novel cell culture substrate comprised of silicon nitride bridges whose structure stiffness can be regulated by altering the axial residual stress without changing material and geometry properties. Both theoretical calculations and finite element simulations were performed to study the influence of residual stress on structure stiffness of bridges. Then multi-positions AFM bending tests were implemented to measure local stiffness of a single micro bridge so as to verify our predictions. NIH/3T3 mouse fibroblast cells were cultured on our substrates to examine the feasibility of the substrate application for investigating cellular response to microenvironment with variable stiffness. The results showed that cells on the edge region near bridge ends were more spread, elongated and better aligned along the bridge axial direction than those on the bridge center region. The results suggest that cells can sense and respond to the differences of stiffness and stiffness gradient between the edge and the center region of the bridges, which makes this kind of substrates can be applied in some biomedical fields, such as cell migration and wound healing.