Bioassembly of multicellular spheroids to mimic complex tissue structure using surface-modified magnetized nanofibers

Abstract

Advancements in biofabrication have led to major strides toward creating authentic organ models; however, replicating intricate organ structures without scaffolds remains challenging. In this study, we introduce a method utilizing surface-modifiable magnetic nanofibers to achieve precise control over spheroid functions and geometrical features, allowing the creation of multiple functional domains within a single microtissue. We generated magnetized nanofibers by electrospinning magnetic nanoparticles dispersed in poly-L-lactic acid solution. These fibers were then coated with polydopamine (PD) to enhance their biological functions, particularly reactive oxygen species (ROS) scavenging. These PD-coated magnetic fibers (PMFs) had magnetic-responsive properties when incorporated into human dermal fibroblast spheroids (0.019 +/- 0.001 emu g-1). Furthermore, PMFs within the spheroids effectively regulated ROS levels by upregulating the expression of key anti-oxidative genes such as superoxide dismutase-1 (2.2 +/- 0.1) and glutathione peroxidase-1 (2.6 +/- 0.1). By exploiting the magnetic responsiveness of spheroids, we were able to assemble them into various structures such as linear, triangular, and square structures using remotely applied magnetic forces. Within the assembled three-dimensional constructs, the cells in spheroids incorporating PMFs demonstrated resistance to ROS regulatory activity in the presence of hydrogen peroxide, while spheroids composed of bare fibers exhibited high ROS levels. Furthermore, we assembled spheroids containing fibroblasts and endothelial cells into complex tissue structures resembling vessels under magnetic manipulation. This innovative method holds tremendous promise for organ modeling and regenerative medicine due to the unprecedented control it allows in developing microtissues that closely emulate real organs.