Multi-Photon Polymerization (MPL) is a Direct Laser Writing (DLW) technique that combines ultrafast (femtosecond, fs) laser pulses and Computer Aided Designs (CADs) for the fabrication of high precision scaffolds that find application in fields such as tissue engineering [1, 2]. We used such scaffolds for mono- and co- cultures of murine N2a neuronal and SW10 glial cells in order to investigate how topography affects the cell behavior on the 3D environment for various timepoints.
A novel bridge-shaped 3D designed of dimensions of 400μm x 400μm x 60μm was fabricated using a femtosecond fiber laser operating at 780 nm (pulse duration: 120 fs, repetition rate 80 MHz). The material used for polymerization was a hybrid material consisting of organic and inorganic components (3-Trimethoxysilyl propyl methacrylate, MAPTMS/ Methacrylic acid, MAA and Zirconium propoxide, ZPO). 4,4’- Bis (diethylamino) benzophenon, Bis, and Sudan Black B were both used as photoinitiators (PIs). The fabricated 3D structures were used as scaffolds for the mono- and co-cultures of N2A neuronal and SW10 glial cells for timepoints starting from 7 days. Cultures were monitored both by Scanning Electron Microscopy (SEM) and Confocal Microscopy for both morphological and intra-cellular investigation.
Cell cultures were conducted on both 3D scaffolds and glass coverslips for various timepoints. Cell growth and survival between the different conditions/ culture periods were investigated to determine the optimal culturing conditions. Comparison of the cultures exhibited a strong preference of directionality for cell elongation and axon growth dictated by the topography of the scaffolds compared to the control glass coverslips. Our findings not only show that our scaffolds can sustain both mono and co- cultures of N2a and SW10 cells, but also that by carefully designing a suitable topography, cell behavior can be influenced towards a desired way.
Our findings show the effect of topographical properties on cell growth and behavior and the ability to influence the aforementioned behavior in a beneficial way by designing 3D scaffolds with specific geometries based on the application. We highlight the potential of the development of an in vitro model for the study of neurodegenerative diseases which may find further application in tissue regeneration.
In2Sight: Horizon 2020 GA: 964481
1. Sun, H.B. and Kawata, S., Eds., ed Berlin, Heidelberg: Springer Berlin Heidelberg, 170: 169-273 (2004).
2. Nguyen, A. K. and Narayan, R. J., Materials Today, 20, (6):314-322 (2017).