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Culturing cells in three-dimensional (3D) environments has been shown to significantly influence cell function, and may provide a more physiologically relevant environment within which to study the behavior of specific cell types. 3D tissues typically present a topologically complex fibrous adhesive environment, which is technically challenging to replicate in a controlled manner. Micropatterning technologies have provided significant insights into cell-biomaterial interactions, and can be used to create fiber-like adhesive structures, but are typically limited to flat culture systems; the methods are difficult to apply to topologically-complex surfaces. In this work, we utilize crack formation in multilayered microfabricated materials under applied strain to rapidly generate well-controlled and topologically complex 'fiber-like' adhesive protein patterns, capable of supporting cell culture and controlling cell shape on three-dimensional patterns. We first demonstrate that the features of the generated adhesive environments such as width, spacing and topology can be controlled, and that these factors influence cell morphology. The patterning technique is then applied to examine the influence of fiber structure on the nuclear morphology and actin cytoskeletal structure of cells cultured in a nanofibrous biomaterial matrix.

Defined topologically-complex protein matrices to manipulate cell shape via three-dimensional fiber-like patterns