Background: Walking with an artificial limb is essential to restore mobility and maintain the quality of life of individuals suffering from limb absence. Bespoke clinical decisions are made to maximise successful fitting of the prosthesis. Unfortunately, satisfactory prosthetic attachment is rarely achieved once the residuum health is compromised. Prosthetic care providers such as rehabilitation and pain specialists, prosthetists, physiotherapists, use medical imaging and embedded bio-sensors to diagnose residuum neuromusculoskeletal disfunctions. However, interfacing these devices and merging complementary data is challenging. This technology gap creates needs for diagnostics device capable of conducting patient-specific differential diagnosis of common neuromusculoskeletal disfunctions. Our purpose is to present the next-generation diagnostic device, called the In-vivo Kinetic System 2.0 integrating loading measurements, dynamic anatomical ultrasonography (DAU), and computational models for a real-time personalized animation of digital twin of the residuum during true loading conditions. Method: The In-vivo Kinetic System 2.0 was developed using the Biodesign innovation process. Results: The proposed wearable and non-invasive Invivo Kinetic System 2.0 diagnostic device could holistically establish the cause-effect relationship between prosthetic care interventions and residuum neuromusculoskeletal disfunctions. It could be used safely, efficiently and routinely by qualified clinicians at critical points of care. Discussion: Current developments of holistic diagnostic devices require to overcome significant barriers associated with the design (e.g., loading measurements, topography of residuum tissues during real-life activities), computational modelling, and clinical roll-out (e.g., selection of primary user). Conclusion: Next-generation diagnostic devices should not only support evidence-based patient-specific prosthetic care, patient empowerment and development of bionics solutions but positively disrupt organisation of healthcare by enabling cost-utility analyses and addressing healthcare gap due to labour shortage. Hopefully, the proposed In-vivo Kinetic System 2.0 will contribute to increase comfort, mobility, and quality of life of the growing population of individuals suffering from limb loss worldwide.