Two-Photon Polymerization of Sub-micrometric Patterned Surfaces: Investigation of Cell-Substrate Interactions and Improved Differentiation of Neuron-like Cells

Year: 2013

Authors: Marino A., Ciofani G., Filippeschi C., Pellegrino M., Pellegrini M., Orsini P., Pasqualetti M., Mattoli V., Mazzolai B.

Autors Affiliation: Center for Micro-BioRobotics @SSSA, Istituto Italiano di Tecnologia, Viale Rinaldo Piaggio 34, 56025 Pontedera, Italy; The Biorobotics Institute, Scuola Superiore Sant’Anna, Viale Rinaldo Piaggio 34, 56025 Pontedera, Italy; Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia University of Pisa, Via Savi 10, 56126 Pisa, Italy; Scuola Normale Superiore di Pisa, Piazza dei Cavalieri 7, 56126 Pisa, Italy; Dipartimento di Biologia, University of Pisa, Via Luca Ghini 13, 56126 Pisa, Ital

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-SYSY derived neurons differentiate on parallel 2.5 mu m 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 mu m) 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.


Volume: 5 (24)      Pages from: 13012  to: 13021

KeyWords: Direct Laser Writing; bio/nonbio interfaces; neuronal tissue engineering; nerve regeneration; axonal guidance
DOI: 10.1021/am403895k

Citations: 74
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