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Engineered cartilage tissue is one of the most promising treatments for articular cartilage defects. In this study, a bioreactor was designed to implement a non-invasive real-time monitoring of the neo-cartilage tissue formation processes through ultrasonic signal analysis. Polylactic acid (PLA) scaffolds were printed and seeded with human chondrocytes. Then, they were cultured in an ultrasound (US)-integrated bioreactor. The readings from the ultrasonic sensors were analyzed by numerical models of the ultrasound-tissue interaction and by a stochastic treatment to infer the extracellular matrix (ECM) evolution. To reconstruct the velocity and attenuation from the recorded signals, a genetic-algorithm based inverse problem (IP) was combined with an iterative computational propagation. The ultrasonic data were validated against evolution measurements of the in vitro 3D chondrocyte cultures assessed by proliferation …