| AVS 54th International Symposium | |
| Biomaterial Interfaces | Tuesday Sessions |
| Session BI-TuM |
| Session: | Proteins at Interfaces |
| Presenter: | M. Ramalingam, University of Central Florida |
| Authors: | M. Ramalingam, University of Central Florida S. Kootala, University of Central Florida N. Bhargava, University of Central Florida M. Stancescu, University of Central Florida M. Hirsh-Kuchma, University of Central Florida M. Klimov, University of Central Florida P. Molnar, University of Central Florida J.J. Hickman, University of Central Florida |
| Correspondent: | Click to Email |
Spatial positioning of neurons on patterned surfaces and characterization of their functional synaptic connectivity and specific extra-cellular matrix (ECM) protein productivity is of great importance for the fabrication of neuron-based biosensors and in developmental cell biology. We have determined that a combination of traditional biological analysis techniques, such as SDS-PAGE and PT-PCR, and surface analytical techniques, such as X-ray Photoelectron Spectroscopy (XPS) and SIMS, is a good approach for ECM analysis. Here, we report on ECM deposition on patterns of hippocampal neurons on surfaces composed of self-assembled monolayers (SAMs) of two different organic compounds, trimethoxysilylpropl-diethylenetriamine (DETA) and tridecafluoro-1,1,2,2,-tetrahydroctyl-1-tricholorsilane (13F) as well as unpatterned controls, in a serum-free culture condition. The patterns were characterized by XPS, contact angle goniometry, electroless metallization and surface profilometry to confirm their surface composition, wettability and topography. Immunostaining of cultured neurons for synapsin I and microtubule-associated proteins (MAP-2) confirmed the pre- and post-synaptic formation. The electrophysiological study of neurons cultured for 14 days further confirmed the functional synaptic connectivity. The deposition and composition of ECM proteins were determined by immunocytochemistry, confocal laser spectroscopy and reverse transcriptase –polymerase chain reaction (RT-PCR), and it was found that the neurons produce laminin, collagen, fibronectin and vitronectin at differing amounts depending on the conditions. We have quantified the amounts of these proteins using Western Blot and SIMS spectroscopy. The overall results indicate that the neurons cultured on patterns secrete ECM proteins in a differential fashion and these data will have significant implications in engineering functional neuronal systems and hybrid devices.