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.

Vocabulary learning through use of the picture-word inductive model for young English learners in China: A mixed methods examination using Cognitive Load Theory

English has been taught as a core and compulsory subject in China for decades. Recently, the demand for English in China has increased dramatically. China now has the world's largest English-learning population. The traditional English-teaching method cannot continue to be the only approach because it merely focuses on reading, grammar and translation, which cannot meet English learners and users' needs (i.e., communicative competence and skills in speaking and writing). ^ This study was conducted to investigate if the Picture-Word Inductive Model (PWIM), a new pedagogical method using pictures and inductive thinking, would benefit English learners in China in terms of potential higher output in speaking and writing. With the gauge of Cognitive Load Theory (CLT), specifically, its redundancy effect, I investigated whether processing words and a picture concurrently would present a cognitive overload for English learners in China. ^ I conducted a mixed methods research study. A quasi-experiment (pretest, intervention for seven weeks, and posttest) was conducted using 234 students in four groups in Lianyungang, China (58 fourth graders and 57 seventh graders as an experimental group with PWIM and 59 fourth graders and 60 seventh graders as a control group with the traditional method). No significant difference in the effects of PWIM was found on vocabulary acquisition based on grade levels. Observations, questionnaires with open-ended questions, and interviews were deployed to answer the three remaining research questions. A few students felt cognitively overloaded when they encountered too many writing samples, too many new words at one time, repeated words, mismatches between words and pictures, and so on. Many students listed and exemplified numerous strengths of PWIM, but a few mentioned weaknesses of PWIM. The students expressed the idea that PWIM had a positive effect on their English teaching. ^ As integrated inferences, qualitative findings were used to explain the quantitative results that there were no significant differences of the effects of the PWIM between the experimental and control groups in both grade levels, from four contextual aspects: time constraints on PWIM implementation, teachers' resistance, how to use PWIM and PWIM implemented in a classroom over 55 students.^

Vocabulary Learning through Use of the Picture-Word Inductive Model for Young English Learners in China: A Mixed Methods Examination Using Cognitive Load Theory

English has been taught as a core and compulsory subject in China for decades. Recently, the demand for English in China has increased dramatically. China now has the world’s largest English-learning population. The traditional English-teaching method cannot continue to be the only approach because it merely focuses on reading, grammar and translation, which cannot meet English learners and users’ needs (i.e., communicative competence and skills in speaking and writing). This study was conducted to investigate if the Picture-Word Inductive Model (PWIM), a new pedagogical method using pictures and inductive thinking, would benefit English learners in China in terms of potential higher output in speaking and writing. With the gauge of Cognitive Load Theory (CLT), specifically, its redundancy effect, I investigated whether processing words and a picture concurrently would present a cognitive overload for English learners in China. I conducted a mixed methods research study. A quasi-experiment (pretest, intervention for seven weeks, and posttest) was conducted using 234 students in four groups in Lianyungang, China (58 fourth graders and 57 seventh graders as an experimental group with PWIM and 59 fourth graders and 60 seventh graders as a control group with the traditional method). No significant difference in the effects of PWIM was found on vocabulary acquisition based on grade levels. Observations, questionnaires with open-ended questions, and interviews were deployed to answer the three remaining research questions. A few students felt cognitively overloaded when they encountered too many writing samples, too many new words at one time, repeated words, mismatches between words and pictures, and so on. Many students listed and exemplified numerous strengths of PWIM, but a few mentioned weaknesses of PWIM. The students expressed the idea that PWIM had a positive effect on their English teaching. As integrated inferences, qualitative findings were used to explain the quantitative results that there were no significant differences of the effects of the PWIM between the experimental and control groups in both grade levels, from four contextual aspects: time constraints on PWIM implementation, teachers’ resistance, how to use PWIM and PWIM implemented in a classroom over 55 students.