Chitin-based anisotropic nanostructures of butterfly wings for regulating cells orientation

In recent years, multiple types of substrates have been applied for regulating cell orientation. Among them, surface topography patterns with grooves or ridges have been widely utilizing for cell culturing. However, this construction is still complicated, low cost-effective and exhibits some technological limitations with either “top-down” or “bottom-up” approaches. Here, a simple and green method was developed by utilizing butterfly wings (Morpho menelaus, Papilio ulysses telegonus and Ornithoptera croesus lydius) with natural anisotropic nanostructures to generate cell alignment. A two-step chemical treatment was proposed to achieve more hydrophilic butterfly wings preceding cell culturing. Furthermore, calcein acetoxymethyl ester (Calcein-AM) staining and Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay results demonstrated the appropriate viability of NIH-3T3 fibroblast cells on those butterfly wings. Moreover, the cells displayed a high degree of alignment in each specimen of these wings. We anticipate that those originating from natural butterfly wings will pose important applications for tissue engineering.