title: Lithography-free fabrication of reconfigurable substrate topography for contact guidance.
abstract: Mammalian cells detect and respond to topographical cues presented in natural and synthetic biomaterials both in vivo and in vitro. Micro- and nano-structures influence the adhesion, morphology, proliferation, migration, and differentiation of many phenotypes. Although the mechanisms that underpin cell-topography interactions remain elusive, synthetic substrates with well-defined micro- and nano-structures are important tools to elucidate the origin of these responses. Substrates with reconfigurable topography are desirable because programmable cues can be harmonized with dynamic cellular responses. Here we present a lithography-free fabrication technique that can reversibly present topographical cues using an actuation mechanism that minimizes the confounding effects of applied stimuli. This method utilizes strain-induced buckling instabilities in bilayer substrate materials with rigid uniform silicon oxide membranes that are thermally deposited on elastomeric substrates. The resulting surfaces are capable of reversible of substrates between three distinct states: flat substrates (A = 1.53 ± 0.55 nm; Rms = 0.317 ± 0.048 nm); parallel wavy grating arrays (A∥= 483.6 ± 7.8 nm; λ∥= 4.78 ± 0.16 μm); perpendicular wavy grating arrays (A⊥= 429.3 ± 5.8 nm; λ⊥= 4.95 ± 0.36 μm). The cytoskeleton dynamics of 3T3 fibroblasts in response to these surfaces was measured using optical microscopy. Fibroblasts cultured on dynamic substrates that are switched from flat to topographic features (FLAT-WAVY) exhibit a robust and rapid change in gross morphology as measured by a reduction in circularity from 0.30 ± 0.13 to 0.15 ± 0.08 after 5 min. Conversely, dynamic substrate sequences of FLAT-WAVY-FLAT do not significantly alter the gross steady-state morphology. Taken together, substrates that present topographic structures reversibly can elucidate dynamic aspects of cell-topography interactions.
authors.affiliation: Department of Biomedical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh 15213, PA, USA.
authors.forename: Pitirat
authors.initials: P
authors.lastname: Pholpabu
authors.affiliation: Department of Biomedical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh 15213, PA, USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: Kustra
authors.affiliation: Department of Materials Science and Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh 15213, PA, USA.
authors.forename: Haosheng
authors.initials: H
authors.lastname: Wu
authors.affiliation: Department of Materials Science and Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh 15213, PA, USA.
authors.forename: Aditya
authors.initials: A
authors.lastname: Balasubramanian
authors.affiliation: Department of Biomedical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh 15213, PA, USA; Department of Materials Science and Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh 15213, PA, USA. Electronic address: cjbetti@gmail.com.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Bettinger
mesh_terms: D041721:3T3-L1 Cells; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D051379:Mice; D013499:Surface Properties
chemical_list: D001672:Biocompatible Materials
keywords: Contact guidance; Microfabrication; Microstructure; Topography