M.S. Thesis Presentation by Russell E. Watts
Tuesday, July 10, 2001

(Drs. Robert Cargill and Andres Garcia, co-advisors)

"Evaluation of DETA as a Surface Treatment to Enhance Neuronal Attachment to a Silicone-Based Substrate"

Abstract

In vitro models of neuronal response to mechanical injury play a large role in discerning the biological response in TBI as well as for evaluating the efficacy of treatments.  However, a substrate is necessary which will support both neuronal culture and the application of mechanical deformation.  A silicone elastomer (Sylgard) treated with 3-[2-(2-aminoethylamino)ethylamino]- propyltrimethoxysilane (DETA) is evaluated as a potential culture substrate for primary neurons.

DETA has been shown to increase neuronal adherence on glass and silicon substrates.  Sylgard treated with DETA was compared to Sylgard with adsorbed poly-L-lysine (PL), Sylgard treated with the combination, and untreated Sylgard. DETA treatment alone or in combination with poly-L-lysine was found to enhance neuronal adhesion and growth. Cells on the DETA-treated Sylgard displayed a more typical neuronal morphology (similar to neurons on tissue culture-treated plastic), had a stronger attachment than did cells cultured on poly-L-lysine, and showed a definite response to stretch injury.

Reasons to explain the difference in attachment were explored as well.  Hydrophilicity does not explain the difference, as determined by goniometer experiments.  Indirect immunocytochemistry does reveal that certain extra-cellular matrix components may be responsible for the difference in attachment.