PC12 transformation to neuron cells activated by low-level laser at 660 nm on the UV pre-treated CR-39 scaffolds with parallel microchannels

Abstract

A rapid method of neuron regeneration is introduced on a novel synthetic scaffold. Initially, poly allyldiglycol carbonate (CR-39) substrate is textured under high dose of ArF UV excimer laser (1000 shot, 300 mJ/pulse at 193 nm) to create superficial periodic parallel microchannels with micrometer spacing and sub-micron width. Ultra violet treated CR-39 (UT CR-39) provides a suitable scaffold to speed up the transformation/differentiation of PC12 cells. The latter is pheochromocytoma from the rat adrenal medulla as an embryonic origin from the neural crest usually exposed to the nerve growth factor (NGF). In fact, PC12 cells are seeded on the microchannels and simultaneously stimulated by coherent red photons at 660 nm within the therapeutic window. The parallel microchannels improve oxygen and water diffusion in the scaffold providing better condition for the cell culture alongside the axon alignment to support the nerve outgrowth. The surface cross-linking due to UV exposure and subsequent induced hydrophilicity notably contribute in the neuron cell regeneration without adding NGF. The cell activation due to the coherent photons give rise to enhance the regeneration process too. Here, we have shown that PC12 cells are efficiently transformed to nerves according to Immunocytochemistry (ICC) and Western Blot verification tests based on MAP2 and Synapsin1 protein antibodies. In general, UT CR-39 acts as a superior ridged bed to elevate the population of neural cells more than three times against those of untreated (control ones). Furthermore, the UT CR-39 scaffold undergoes extra improvement of ~ 30% after 12 minutes laser activation regarding the photo biomodulation (PBM) mechanism.