Humanoid Robotic Loading Enhances Mechanotransduction in Tendon Tissue Engineering

Mechanical stimulation is essential in tissue engineering and regenerative medicine for proper tissue maturation. However, conventional uniaxial platforms fail to reproduce the multiaxial loading experienced in vivo. In this study, we present a humanoid robotic bioreactor capable of delivering human-like shoulder motions to engineered tendon constructs, enabling controlled multiaxial stimulation with real-time strain monitoring. Human mesenchymal stem cells were cultured on decellularized tendon scaffolds and subjected to adduction–abduction loading at peak strains of approximately 3.5% and 9.5% under external forces of 25 and 50 N, respectively. Strain levels were directly quantified in situ using a flexible sensor integrated within the bioreactor. The transparent bioreactor membrane allowed noninvasive observation while simultaneously applying mechanical stimulation over 14 d, with continuous assessment of cellular morphology without fixation. Compared with static and traditional uniaxial controls, the robot motions enhance cell alignment and activation of mechanotransduction pathways while inducing notable gene and protein expression changes, particularly within the PI3K–Akt signaling pathway. Although dynamic loading resulted in a moderate reduction in cell viability, the transcriptional profile was consistent with mechanically driven phenotypic adaptation toward tenogenic-related programs rather than dominant signatures of acute cytotoxic damage. These findings demonstrate that replicating human-like multiaxial mechanics in vitro fundamentally alters cellular mechanosensing and may provide a mechanobiological foundation for the future development of more physiologically relevant tendon grafts.