title: Two-photon polymerization of sub-micrometric patterned surfaces: investigation of cell-substrate interactions and improved differentiation of neuron-like cells.
abstract: Direct Laser Writing (DLW) is an innovative tool that allows the photofabrication of high resolution 3D structures, which can be successfully exploited for the study of the physical interactions between cells and substrates. In this work, we focused our attention on the topographical effects of submicrometric patterned surfaces fabricated via DLW on neuronal cell behavior. In particular, we designed, prepared, and characterized substrates based on aligned ridges for the promotion of axonal outgrowth and guidance. We demonstrated that both rat PC12 neuron-like cells and human SH-SY5Y derived neurons differentiate on parallel 2.5 μm spaced submicrometric ridges, being characterized by strongly aligned and significantly longer neurites with respect to those differentiated on flat control substrates, or on more spaced (5 and 10 μm) ridges. Furthermore, we detected an increased molecular differentiation toward neurons of the SH-SY5Y cells when grown on the submicrometric patterned substrates. Finally, we observed that the axons can exert forces able of bending the ridges, and we indirectly estimated the order of magnitude of these forces thanks to scanning probe techniques. Collectively, we showed as submicrometric structures fabricated by DLW can be used as a useful tool for the study of the axon mechanobiology.
authors.affiliation: Center for Micro-BioRobotics @SSSA, Istituto Italiano di Tecnologia, Viale Rinaldo Piaggio 34, 56025 Pontedera, Italy.
authors.forename: Attilio
authors.initials: A
authors.lastname: Marino
authors.forename: Gianni
authors.initials: G
authors.lastname: Ciofani
authors.forename: Carlo
authors.initials: C
authors.lastname: Filippeschi
authors.forename: Mario
authors.initials: M
authors.lastname: Pellegrino
authors.forename: Monica
authors.initials: M
authors.lastname: Pellegrini
authors.forename: Paolo
authors.initials: P
authors.lastname: Orsini
authors.forename: Massimo
authors.initials: M
authors.lastname: Pasqualetti
authors.forename: Virgilio
authors.initials: V
authors.lastname: Mattoli
authors.forename: Barbara
authors.initials: B
authors.lastname: Mazzolai
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D006801:Humans; D007834:Lasers; D009474:Neurons / Q000166:cytology*; D016716:PC12 Cells; D017785:Photons; D058105:Polymerization; D051381:Rats; D013499:Surface Properties; D023822:Tissue Engineering