Crafting Precision: Design and Fabrication of a Xurography-Driven Microfluidic Platform for Exploring Neuron Culture and Targeted Drug Screening.

Abstract

Over the past few decades, rapid advancements among in vitro cell culture models have come up as valuable assets in neurobiological research to understand the complexities of the human brain, disease development and progression at cellular and molecular levels. However, conventional 2D cell culture methods often fail to provide deeper insights into the complex phenomenon of neural cytoarchitecture. This limitation has led to the development of neural organoids such as neurospheres, which offer a closer representation of several neuronal cells. Microfluidics-based neuronal culture platforms further enhance neurosphere generation by enabling precise spatiotemporal control of physical and chemical cues. Here we report the design and fabrication of a low-cost, novel microfluidic device using a cutting-edge and cost-effective xurography technique. We further performed primary neuron culture, forming neurospheres and single cells at varying seeding densities inside the microchannels. Furthermore, to validate the compatibility of the microfluidic device for neuronal disease model generation, we cultured SH-SY5Y cell lines and checked their differentiation inside the chamber. Additionally, we demonstrated the application of the fabricated device as a coculture model using astrocytes and neurons. Finally, in an Alzheimer's disease model context, we tested the device using a multitargeted compound, TDSB, with three important moieties to manage reactive oxygen species in the differentiated SH-SY5Y cells. The results revealed that TDSB can decrease metal-induced ROS generation and inhibit Aβ-Cu(II) induced cytotoxicity. Therefore, our multifaceted microfluidic device can open avenues for neuronal culture and coculture, neurodegenerative disease modeling, and screening of novel neurotherapeutic interventions.