Abstract
In this study, three-dimensional (3D) multi-component microstructures were precisely fabricated via multiphoton excited photochemistry using a femtosecond laser direct-writing system with proposed repetition positioning and vector scanning techniques. Extracellular matrix (ECM) proteins, such as fibronectin (FN), are difficult to stack and form 3D structures larger than several-hundred microns in height due to the nature of their protein structure. Herein, to fabricate complex 3D microstructures with FN, a 3D scaffold was designed and formed from bovine serum albumin (BSA), after which human FN was inserted at specific locations on the BSA scaffold; in this manner, the fabricated ECM microstructure can guide cells in a 3D environment. A human breast cancer cell line, MDA-MB-231, was used to investigate the behavior of cell migration and adhesion on the fabricated human FN and BSA protein structures. Experimental results indicate that many cells are not able to attach or climb on a 3D structure's inclined plane without FN support; hence, the influence of cell growth in a 3D context with FN should being taken into consideration. This 3D multi-protein fabrication technique holds potential for cell studies in designed complex 3D ECM scaffolds.
Highlights
Complex components of extracellular matrix (ECM) proteins play important roles in cell biology, including wound-healing and tumor metastasis
Fibrous collagen or matrigel matrices are commonly used for the study of 3D cell behavior; these matrices have random pore sizes and are structurally and chemically ill-defined, which results in discrepancies between cell behavior in vivo and in artificial two-dimensional (2D) environments
It should be noted that the bovine serum albumin (BSA) monolayer can be used as a nonspecific surface for comparison with the adhesion dynamics of the cells on or off the crosslinked ECM protein structures
Summary
Complex components of extracellular matrix (ECM) proteins play important roles in cell biology, including wound-healing and tumor metastasis They may form niche circumstances of tissue engineering scaffolds, since these can influence cell adhesion and migration behavior in three-dimensional (3D) surroundings and promote matrix synthesis. There are many different configurations for fabricating complex 3D microstructures, which can be roughly classified into two types: mask projection and point-wise scan With respect to the former, several mask projection configurations have been demonstrated that can fabricate 3D scaffolds via a digital micromirror device or optical mask [8,9,10]; these techniques have poor axial resolution for 3D fabrication due to the unfilled back-focal aperture. The proposed 3D multi-protein fabrication technique with its unique capability of precisely processing and manufacturing of large-scale, multi-component, and elegant freeform ECM protein structures could offer great potential in the field of 3D tissue engineering and in vitro biomedical research for simulating in vivo environments
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