研究目的
To establish the relationship between the micro/nano structures of the P3HT substrates and the neuronal response, and to engineer semiconducting polymers towards noninvasive photostimulation of neural cells and tissues.
研究成果
P3HT-based substrates with micro/nano structures enable wireless photostimulation, enhancing neuronal differentiation and growth. Nanofibers promote more branches, while microfibers lead to longer neurites. This approach shows promise for neural engineering and regenerative devices, with future work needed for in vivo applications and detailed mechanistic studies.
研究不足
The study is limited to in vitro experiments with PC12 cells; potential issues include cell migration across substrate regions during culturing and irradiation, and the need for further in vivo validation. The photoconductive effect may be influenced by substrate dimensions and material properties.
1：Experimental Design and Method Selection:
The study involved fabricating P3HT-based substrates with different morphologies (homogeneous films, self-assembled nanofibers, electrospun microfibers, and lithographically patterned stripes) to investigate their photoconductive effects on neuronal cells under light irradiation. Methods included self-assembly, electrospinning, photolithography, and cell culture techniques.
2：Sample Selection and Data Sources:
Rat pheochromocytoma (PC12) cells were used as a model for primary neurons. Substrates were prepared with specific dimensions: nanofibers (~100 nm diameter), microfibers (~1 μm diameter), and patterned stripes (3, 25, 50 μm width).
3：List of Experimental Equipment and Materials:
Equipment included AFM, SEM, TEM, laser confocal microscopy, optical microscopy, LED light source (2 W power), and electrospinning setup. Materials included P3HT, anisole, chloroform, poly(ε-caprolactone) (PCL), poly-D-lysine, Fluo-4 AM dye, and Alexa Fluor? 633 phalloidin.
4：Experimental Procedures and Operational Workflow:
Substrates were fabricated (e.g., spin-coating for films, slow cooling for nanofibers, electrospinning for microfibers, photolithography for patterns), treated with poly-D-lysine for cell adhesion, seeded with PC12 cells, irradiated with green LED light, and analyzed using microscopy and staining techniques over 3 days.
5：Data Analysis Methods:
Neurite extensions were measured and statistically analyzed (e.g., average lengths, Gaussian fits) from microscopy images; intracellular calcium changes were monitored using Fluo-4 AM fluorescence.
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