Nanovibrational stimulation preferentially enhances osteogenic responses in zebrafish

Abstract Mechanical cues are key regulators of bone formation, yet their potential as therapeutic stimuli remains incompletely explored in vivo. Nanovibrational stimulation, which delivers low-amplitude, high-frequency mechanical input, has been shown to promote osteogenic differentiation in vitro, and rodent studies have similarly demonstrated osteogenic effects. However, its impact on other cellular systems at the whole-organism level remains poorly understood. Here, we demonstrate that nanovibrational stimulation enhances myogenic differentiation in vitro before investigating its effects on skeletal development and tissue specificity in zebrafish. Larval zebrafish exposed to nanovibrational stimulation exhibited increased osteoblast numbers and enhanced bone formation relative to controls. In adult zebrafish, nanovibration increased the osteoblast response to a fracture-like injury, indicating enhanced osteogenic activity during repair. To assess tissue specificity, we examined additional cell types and systems relevant to skeletal regeneration, including cartilage, muscle, vasculature and innate immune cells. Although nanovibrational stimulation promoted myogenic differentiation of C2C12 cells in vitro, its effects on muscle and other non-skeletal tissues in zebrafish larvae were comparatively limited. These findings suggest that nanovibrational stimulation exerts a preferential effect on osteogenic processes in vivo. These findings demonstrate that nanovibrational stimulation preferentially enhances osteoblast-mediated bone formation and skeletal injury responses in zebrafish, without causing significant perturbations in other tissues. Our results establish zebrafish as a tractable in vivo model for investigating vibration-induced mechanobiological processes, providing a more physiologically relevant representation of tissue-level mechanotransduction than conventional two-dimensional culture systems. Furthermore, these findings highlight the potential of nanovibrational stimulation as a non-invasive strategy to promote bone regeneration and fracture repair.