This study aims to enhance the understanding of the relationship between Ilizarov fixator configuration and its effects on bone fracture healing. Using Taylor spatial frame (TSF) as an example, the roles of critical parameters (ie, TSF ring diameter, wire pre-tension, fracture gap size, and axial load) that govern fracture healing during the early stages were investigated by using computational modelling in conjunction with mechanical testing involving an advanced 3D optical measurement system. The computational model was first validated using the mechanical test results and then used to simulate mesenchymal stem cell (MSC) differentiations within different regions of the fracture site under various combinations of TSF ring diameter, wire pre-tension, fracture gap size, and axial load values. Predicted spatially dependent MSC differentiation patterns and the influence of each parameter on differentiations were compared with in vivo results, and good agreement was seen between the two. Gap size was identified as the most influential parameter in MSC differentiation, and the influence of axial loading and TSF configuration (ie, ring diameter and wire pre-tension) on cell differentiation was seen to be gap size dependent. Most changes in cell differentiation were predicted in the external callus (periosteal), which is the crucial region of the callus in the early stages.However, for small gap sizes (eg, 1 mm), significant changes were predicted in the endosteal callus as well. The study exhibits the potential of computational models in assessing the performance of Ilizarov fixators as well as assisting surgeons in patient-specific clinical treatment planning. KEYWORDS 3D optical measurement system, mechanical test, mechano-regulation, mesenchymal stem cell, Taylor spatial frame