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Manufacturing micropatterned collagen scaffolds with chemical-crosslinking for development of biomimetic tissue-engineered oral mucosa

The junction between the epithelium and the underlying connective tissue undulates, constituting of rete ridges, which lack currently available soft tissue constructs. In this study, using a micro electro mechanical systems process and soft lithography, fifteen negative molds, with different dimensions and aspect ratios in grid- and pillar-type configurations, were designed and fabricated to create three-dimensional micropatterns and replicated onto fish-scale type I collagen scaffolds treated with chemical crosslinking. Image analyses showed the micropatterns were well-transferred onto the scaffold surfaces, showing the versatility of our manufacturing system. With the help of rheological test, the collagen scaffold manufactured in this study was confirmed to be an ideal gel and have visco-elastic features. As compared with our previous study, its mechanical and handling properties were improved by chemical cross-linking, which is beneficial for grafting and suturing into the complex structures of oral cavity. Histologic evaluation of a tissue-engineered oral mucosa showed the topographical microstructures of grid-type were well-preserved, rather than pillar-type, a well-stratified epithelial layer was regenerated on all scaffolds and the epithelial rete ridge-like structure was developed. As this three-dimensional microstructure is valuable for maintaining epithelial integrity, our micropatterned collagen scaffolds can be used not only intraorally but extraorally as a graft material for human use.1



Te scafolds include several combinations of four prototypes, with diferent feature dimensions and aspect ratios, as well as presence and absence of EDC crosslinking (1–4). G–R, G–T, P–R, and P–T represent for grid pattern with rectangular confguration, grid pattern with truncated confguration, pillar pattern with rectangular confguration, and pillar patterns with truncated confguration, respectively. Schematic illustration at the bottom of right side shows a basic micropattern design referred in the table within this fgure [(a) height, (b) thickness, (c) channel width].
A total of 16 micropatterns of collagen scafolds designed in this study (C, 1–15)



1. Suzuki, A., Kodama, Y., Miwa, K. et al. Manufacturing micropatterned collagen scaffolds with chemical-crosslinking for development of biomimetic tissue-engineered oral mucosa. Sci Rep10, 22192 (2020). https://doi.org/10.1038/s41598-020-79114-3