Traumatic brain injury (TBI) results from a physical insult to the head and often results in temporary or permanent brain dysfunction;​ however, the cellular pathology remains poorly understood and there are currently no clinically effective treatments. Properly designed and characterized laboratory models that adequately recapitulate the biomechanical and cellular pathophysiology of neural trauma provide valuable experimental platforms for the elucidation of the cellular response to injury as well as serving as test-beds for potential therapeutic interventions. A primary goal of this work was to develop and characterize a novel three-dimensional (3-D) in vitro paradigm of neural trauma integrating a robust 3-D neural co-culture system and a well-defined biomechanical input representative of clinical TBI. Mechanically injured 3-D co-cultures were then interfaced with neural stem cells (NSCs) to serve as an in vitro test-bed for the evaluation of factors influencing NSC survival and integration.

Specifically, a novel 3-D neuronal-astrocytic co-cultures system was developed, establishing parameters resulting in the growth and vitality of mature 3-D networks, potentially providing enhanced physiological relevance and providing a robust platform for the mechanistic study of neurobiological phenomena. Furthermore, an electromechanical device was developed and characterized that is capable of subjecting 3-D cell-containing matrices to a defined mechanical insult, with a predicted strain manifestation at the cellular level - the first model to date capable of correlating cellular outcome with specific 3-D biomechanics. Following independent development and validation, these novel 3-D neural cell and mechanical trauma paradigms were used in combination to develop a mechanically-induced model of neural degeneration and reactive astrogliosis. This vitro surrogate model of a degenerating and reactive astrogliotic environment was then exploited to assess factors influencing NSC fate upon delivery to this environment, revealing that specific factors in a degenerating environment were detrimental to NSC survival. This work has developed enabling technologies for the in vitro study of neurobiological phenomena and responses to neural injury, and may aid in elucidating the complex biochemical and molecular cascades that occur after a traumatic insult. Furthermore, the novel paradigm here developed may provide a powerful experimental framework for improving treatment strategies for therapeutic implementation following CNS injury, and therefore serve as a valid preanimal test-bed.