Double-Orthogonal Gradient-Based High-Throughput Screening Platform for Studying Cell Response Toward Combined Physicochemical Biomaterial Properties

Implant-associated complications arise due to non-optimized cell–biomaterial interactions. It is well known that cells respond to their physicochemical microenvironment on 2D interfaces and 3D networks. Attempts to manipulate this interaction target surface parameters such as wettability (W), stiffness (S), and topography (T) to influence cell differentiation, adhesion, and morphology, due to induction of gene activation and protein expression. Investigating the combinatorial influence of all three mentioned parameters simultaneously remains challenging, though most realistic, since all three parameters are inherently present on a surface. Herein, a novel high-throughput screening technology, which allows investigating the cell response of human bone-marrow-derived mesenchymal stem cells toward three varying biomaterial surface parameters simultaneously, is presented. The platform provides efficient screening and cell response readout to a vast amount of combined biomaterial surface properties, in a single-cell experiment. Surface gradients of aligned wrinkle T, S, and W are orthogonally combined giving four combinatorial surfaces. The screening outcome is validated by translating interesting regions to homogeneous surfaces. Cells are found to behave similar to the screening in terms of adhesion, spreading, and vimentin expression. The technology tremendously supports the identification of optimal surface parameter combinations and potentially addressing many of the current implant-associated complications.