Piezoelectric nanocomposite bioink and ultrasound stimulation modulate early skeletal myogenesis

Despite the significant progress in bioprinting for skeletal muscle tissue engineering, new stimuli-responsive bioinks to boost the myogenesis process are highly desirable. In this work, we developed a printable alginate/Pluronic-based bioink including piezoelectric barium titanate nanoparticles (nominal diameter: similar to 60 nm) for the 3D bioprinting of muscle cell-laden hydrogels. The aim was to investigate the effects of the combination of piezoelectric nanoparticles with ultrasound stimulation on early myogenic differentiation of the printed structures. After the characterization of nanoparticles and bioinks, viability tests were carried out to investigate three nanoparticle concentrations (100, 250, and 500 mu g mL(-1)) within the printed structures. An excellent cytocompatibility was confirmed for nanoparticle concentrations up to 250 mu g mL(-1). TEM imaging demonstrated the internalization of BTNPs in intracellular vesicles. The combination of piezoelectric nanoparticles and ultrasound stimulation upregulated the expression of MYOD1, MYOG, and MYH2 and enhanced cell aggregation, which is a crucial step for myoblast fusion, and the presence of MYOG in the nuclei. These results suggest that the direct piezoelectric effect induced by ultrasound on the internalized piezoelectric nanoparticles boosts myogenesis in its early phases.

Automated summary

By combining piezoelectric nanoparticles with ultrasound stimulation, early myogenesis in muscle cell-laden hydrogels can be enhanced in 3D bioprinting, promoting cell aggregation and crucial steps for myoblast fusion.