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  • Programmed Topographic Substrates for Studying Roughness Gradient-Dependent Cell Migration Using Two-Photon Polymerization.

Programmed Topographic Substrates for Studying Roughness Gradient-Dependent Cell Migration Using Two-Photon Polymerization.

Frontiers in cell and developmental biology (2022-04-09)
Subhashree Shivani, Yu-Hsiang Hsu, Cheng-Je Lee, Chi-Sheng Cheong, Tien-Tung Chung, An-Bang Wang
초록

The mediation of the extracellular matrix is one of the major environmental cues to direct cell migration, such as stiffness-dependent durotaxis and adhesiveness-dependent haptotaxis. In this study, we explore another possible contact guidance: roughness dependent topotaxis. Different from previously reported studies on topotaxis that use standard photolithography to create micron or submicron structures that have identical height and different spatial densities, we develop a new method to programmatically fabricate substrates with different patterns of surface roughness using two-photon polymerization. Surface roughness ranging from 0.29 to 1.11 μm can be created by controlling the voxel distance between adjacently cured ellipsoid voxels. Patterned Ormocomp® masters are transferred to polypropylene films using the nanoimprinting method for cell migration study. Our experimental results suggest that MG63 cells can sense the spatial distribution of their underlying extracellar roughness and modulate their migration velocity and direction. Three characteristic behaviors were identified. First, cells have a higher migration velocity on substrates with higher roughness. Second, cells preferred to migrate from regions of higher roughness to lower roughness, and their migration velocity also decreased with descending roughness. Third, the migration velocity remained unchanged on the lower roughness range on a graded substrate with a steeper roughness. The last cell migration characteristic suggests the steepness of the roughness gradient can be another environmental cue in addition to surface roughness. Finally, the combination of two-photon polymerization and nanoimprint methods could become a new fabrication methodology to create better 3D intricate structures for exploring topotactic cell migrations.

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Sigma-Aldrich
Anti-Vinculin−FITC antibody, Mouse monoclonal, clone hVIN-1, purified from hybridoma cell culture