Date of Award


Document Type



Santa Clara University

First Advisor

Prashanth Asuri

Second Advisor

Christelle Sabatier


Culturing neurons in vitro is a challenging task because they are a highly specialized cell type that reside in a complex and unique environment in the body. The aim of the research presented in the following thesis was to design a biomimetic, three-dimensional scaffold capable of (1) promoting primary neuron maturation and axonal outgrowth and (2) serving as a system for toxicology screening. In the system presented here, neurons were cultured in three-dimensional hydrogels, simulating the physiological environment that these cells experience within the body. In doing so, a biologically relevant response was elicited upon their exposure to acrylamide, a known neurotoxin. Primary neurons were isolated from embryonic chick spinal cords and seeded in alginate or collagen hydrogels. Initial cell viability assays and qualitative microscopy observations confirmed that primary neurons not only survive in the system, but also exhibit positive axon growth. Next, neuronal response to acrylamide was evaluated through cytotoxicity assays and axon outgrowth measurement through immunocytochemistry. The study of neurotoxic effects on neuron maturation in collagen gels yielded two key observations: (1) the level of cell death in neurons exposed to acrylamide is dependent on the density and availability of attachment proteins in the collagen hydrogel environment and (2) the axonal retraction response is more sensitive to acrylamide than the lethal cellular response is. The ability of the collagen scaffold to provide insight into neuronal response to toxin in a biomimetic, three-dimensional environment suggests the system’s potential for increasing the accuracy of early stage cytotoxicity testing for safe and efficient pharmaceutical candidates. Additionally, this research has further implications in neurodegenerative disease modeling, spinal cord injury recovery studies, and basic research in neuroscience and neural tissue engineering.