Three-Dimensional Printing of Pure Proteinaceous Microstructures by Femtosecond Laser Multiphoton Cross-Linking

Laser direct write (LDW) is a promising three-dimensional (3D) printing technology for creating proteinaceous microstructures in which the proteins retain their original function, enabling the manufacture of complex biomimetic 3D microenvironments and versatile enhancement of medical microdevices. A photoactivator has commonly been used to date in the laser direct write of proteins to enhance the cross-linking process. However, incomplete conversion results in photoactivator molecules remaining trapped inside the protein microstructure, causing their gradual leaching and subsequent undesirable effect on biological applications. Here, we demonstrate the 3D fabrication of microstructures made of pure serum albumin protein using photoactivator-free fabrication, confirmed by Raman data. For the first time, acid-catalyzed hydrolysis of the created structures provides evidence that chemical cross-links are induced by exposure to femtosecond laser irradiation. The diversity of the biomaterial protein available for the precursors for LDW offers capability of the fabrication of complex biomimetic 3D microenvironments and biochip applications.