Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)

Introduction: In this study, quasi-three-dimensional (3D) microwell patterns were fabricated with poly (l-lactic acid) for the development of cell-based assays, targeting voltage-gated calcium channels (VGCCs). Methods and materials: SH-SY5Y human neuroblastoma cells were interfaced with the microwell patterns and found to grow as two dimensional (2D), 3D, and near two dimensional (N2D), categorized on the basis of the cells’ location in the pattern. The capability of the microwell patterns to support 3D cell growth was evaluated in terms of the percentage of the cells in each growth category. Cell spreading was analyzed in terms of projection areas under light microscopy. SH-SY5Y cells’ VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium GreenTM-1. The expression of L-type calcium channels was evaluated using immunofluorescence staining with DM-BODIPY. Results: It was found that cells within the microwells, either N2D or 3D, showed more rounded shapes and less projection areas than 2D cells on flat poly (l-lactic acid) substrates. Also, cells in microwells showed a significantly lower VGCC responsiveness than cells on flat substrates, in terms of both response magnitudes and percentages of responsive cells, upon depolarization with 50 mM K+. This lower VGCC responsiveness could not be explained by the difference in L-type calcium channel expression. For the two patterns addressed in this study, N2D cells consistently exhibited an intermediate value of either projection areas or VGCC responsiveness between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness. Conclusion: These results suggest that the pattern structure and therefore the cell growth characteristics were critical factors in determining cell VGCC responsiveness and thus provide an approach for engineering cell functionality in cell-based assay systems and tissue engineering scaffolds.

Optimization of three-dimensional branching networks of microchannels for thermal management of microelectronics

The aim of this work is to present a methodology to develop cost-effective thermal management solutions for microelectronic devices, capable of removing maximum amount of heat and delivering maximally uniform temperature distributions. The topological and geometrical characteristics of multiple-story three-dimensional branching networks of microchannels were developed using multi-objective optimization. A conjugate heat transfer analysis software package and an automatic 3D microchannel network generator were developed and coupled with a modified version of a particle-swarm optimization algorithm with a goal of creating a design tool for 3D networks of optimized coolant flow passages. Numerical algorithms in the conjugate heat transfer solution package include a quasi-ID thermo-fluid solver and a steady heat diffusion solver, which were validated against results from high-fidelity Navier-Stokes equations solver and analytical solutions for basic fluid dynamics test cases. Pareto-optimal solutions demonstrate that thermal loads of up to 500 W/cm2 can be managed with 3D microchannel networks, with pumping power requirements up to 50% lower with respect to currently used high-performance cooling technologies.