Towards understanding the mechanism of 3D printing using protein: Femtosecond laser direct writing of microstructures made from homopeptides

Abstract

Femtosecond laser direct write (fs-LDW) is a promising technology for three-dimensional (3D) printing due to its high resolution, flexibility, and versatility. A protein solution can be used as a precursor to fabricate 3D proteinaceous microstructures that retain the protein's native function. The large diversity of protein molecules with different native functions allows diverse applications of this technology. However, our limited understanding of the mechanism of the printing process restricts the design and generation of 3D microstructures for biomedical applications. Therefore, we used eight commercially available homopeptides as precursors for fs-LDW of 3D structures. Our experimental results show that tyrosine, histidine, glutamic acid, and lysine contribute more to the fabrication process than do proline, threonine, phenylalanine, and alanine. In particular, we show that tyrosine is highly beneficial in the fabrication process. The beneficial effect of the charged amino acids glutamic acid and lysine suggests that the printing mechanism involves ions in addition to the previously proposed radical mechanism. Our results further suggest that the uneven electron density over larger amino acid molecules is key in aiding fs-LDW. The findings presented here will help generate more desired 3D proteinaceous microstructures by modifying protein precursors with beneficial amino acids. STATEMENT OF SIGNIFICANCE: Femtosecond laser direct write (fs-LDW) offers a three-dimensional (3D) printing capability for creating well-defined micro-and nanostructures. Applying this technology to proteins enables the manufacture of complex biomimetic 3D micro-and nanoarchitectures with retention of their original protein functions. To our knowledge, homopeptides themselves have never been used as precursor for fs-LDW so far. Our study gains several new insights into the 3D printing mechanism of pure protein for the first time. We believe that the experimental evidence presented greatly benefits the community of 3D printing of protein in particular and the biomaterial science community in general. With the gained insight, we aspire to expand the possibilities of biomaterial and biomedical applications of this technique.

Keywords: 3D printing; Amino acid; Homopeptide; LDW; Microfabrication; Protein.