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.

Public perception of carbon capture and storage in relation to hydraulic fracturing: content analyses, survey results, and policy proposals

Carbon capture and storage (CCS) can contribute significantly to addressing the global greenhouse gas (GHG) emissions problem. Despite widespread political support, CCS remains unknown to the general public. Public perception researchers have found that, when asked, the public is relatively unfamiliar with CCS yet many individuals voice specific safety concerns regarding the technology. We believe this leads many stakeholders conflate CCS with the better-known and more visible technology hydraulic fracturing (fracking). We support this with content analysis of media coverage, web analytics, and public lobbying records. Furthermore, we present results from a survey of United States residents. This first-of-its-kind survey assessed participants’ knowledge, opinions and support of CCS and fracking technologies. The survey showed that participants had more knowledge of fracking than CCS, and that knowledge of fracking made participants less willing to support CCS projects. Additionally, it showed that participants viewed the two technologies as having similar risks and similar risk intensities. In the CCS stakeholder literature, judgment and decision-making (JDM) frameworks are noticeably absent, and public perception is not discussed using any cognitive biases as a way of understanding or explaining irrational decisions, yet these survey results show evidence of both anchoring bias and the ambiguity effect. Public acceptance of CCS is essential for a national low-carbon future plan. In conclusion, we propose changes in communications and incentives as programs to increase support of CCS.