The Effect of Using Low Intensity Pulse Ultrasound and Culture Substrates Variation on the Regulation of Neural Stem Cells

Recent literature has revealed that neural stem cells (NSCs) have potential for the basic materials for treating harsh neural diseases and injuries, such as Alzheimer's disease and Parkinson's disease. In the proposed research, a regulating microenvromental stimuli system for neural stem cells (NSCs) will be establish including the effect of materials, biophysical stimulation, and biochemical stimulation. Our first goal will be focus on developing a series of microenviroment by using one of a biomimetic material-supported lipid bilayer (SLB) conjugated layer by layer (LBL) polyelectrolyte multilayers (PEM) or ECM proteins as a series template to program and mimic NSCs niches and to determine the cell-cell and cell-substrate interactions. Secondly, one of a biophysical stimulation-low intensity pulse ultrasound (LIPUS) will also be introduced into this system to investigate the proliferation, differentiation, and neuron process outgrowth of NSCs on a series surface stimuli and LIPUS stimulation. During the first year, since SLB formed by the fusion of small unilamellar vesicles on substrate serve as important biomimetic model membranes in both scientific research and practical applications, we will construct a system of PEM or ECM protein conjugated SLB as a molecular model to study the surface properties variation such as charge, thickness, roughness, stiffness, hydrophilic properties, and the mobility of SLB. We will combine sequential measurements by Quartz Crystal Microbalance with Dissipation (QCM-D), atomic force microscopy (AFM) and ellipsometry, to characterize the SLB formation and surface properties variation. The retards of mobility of SLB after PEM or protein conjugation will be estimated by Fluorescence Recovery after Photobleaching (FRAP). In addition, the effect of a series variation of the intensity, stimuli time/day, with different culture condition of LIPUS on the proliferation, differentiation, migration, neurite outgrowth, and the neurons network will also be investigated. The second year, we will culture NSCs on different substrates that have been analyzed in the first year as an in vitro model to simulated the cell-materials interaction for investigation of the NSCs behaviors under different microenviroment. Eventually, the goal of this proposed project is to create a system closer nature physiology. Therefore, the combined effect including materials, biochemical stimulation and LIPUS stimuli will be integrated in the model to demonstrate the effects of complex stimulation of NSCs and try to build up a neural network system which could be used on the application of neural tissue engineering or cell therapy.

Project ID:PB10007-7258
External Project ID:NSC100-2221-E182-006