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Augmentation of alignment and differentiation in C2C12 skeletal myoblasts through use of nano-to-microscale biochemical patterns

Abstract

Interactions between cell surfaces and the extracellular matrix have been shown in previous studies to play an essential role in cell mobility, adhesion, proliferation, differentiation, polarity, and apoptosis. Mimicking this extracellular microenvironment with nanoscale patterns is an approach in which we can manipulate cellular responses at a molecular level for use in future tissue engineering applications or in vitro models. The use of electron beam lithography was explored in this study to create micron and submicron protein patterns in an attempt to imitate the extracellular microenvironment as a cell might sense in an in vivo setting. Submicron patterns of 250 nm and 500 nm widths with 250 nm and 500 nm spacings respectively were successfully generated out of 10k molecular weight polyethylene glycol. The study examined the effects these submicron patterns exhibited over the alignment and differentiation of C2C12 skeletal myoblasts. 5 [Mu]m patterns showed an apparent effect on both C2C12 alignment and early differentiation compared to the unpatterned substrate. Furthermore, the study found that 250 nm patterns were significantly more effective in directing myotube alignment compared to unpatterned and 500 nm patterned substrates. The 500 nm patterns were found to have the least noticeable effects on C2C12 alignment and differentiation. The study has shown that C2C12 myotube alignment can be guided by a variety of pattern dimensions with varying degrees of success. The results also suggest that existence of focal adhesions of sizes 250 nm or less may be responsible for aiding the alignment of myotubes along the patterns

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