Understanding the evolution of callus mechanical properties over time provides insights in the mechanobiology of fracture healing and tissue differentiation, can be used to validate numerical models, and informs clinical practice. Bone transport experiments were performed in sheep, in which a distractor type Ilizarov was implanted. The forces through the fixator evolution were measured and the callus stiffness was estimated from these forces. Computerized tomography images were taken and bone volume of the callus at different stages was obtained. The results showed that the maximum bone tissue production rate (0.146 cm³/day) was achieved 20 days after the end of the distraction phase. 50 days after the end of the distraction phase, the callus was ossified completely and had its maximum volume, 6-10 cm³ In addition, 80-90% of the load sustained by the operated limb was recovered and the callus stiffness increased exponentially until 5.4-11.4 kN/mm, still below 10% of the healthy level of callus stiffness. The effects of the bony bridging of the callus and the time of the fixator removal on callus force, stiffness and volume were analyzed. These outcomes allowed relating quantifiable biological aspects (callus volume and tissue production rate) with mechanical parameters (callus force and stiffness) using data from the same experiment.

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The load bearing characteristics of the intervened limb over time in vivo are important to know in distraction osteogenesis and bone healing for the characterization of the bone maturation process. Gait analyses were performed for a group of sheep in which bone transport was carried out. The ground reaction force was measured by means of a force platform, and the gait parameters (i.e., the peak, the mean vertical ground reaction force and the impulse) were calculated during the stance phase for each limb. The results showed that these gait parameters decreased in the intervened limb and interestingly increased in the other limbs due to the implantation of the fixator. Additionally, during the process, the gait parameters exponentially approached the values for healthy animals. Corresponding radiographies showed an increasing level of ossification in the callus. This study shows, as a preliminary approach to be confirmed with more experiments, that gait analysis could be used as an alternative method to control distraction osteogenesis or bone healing. For example, these analyses could determine the appropriate time to remove the fixator. Furthermore, gait analysis has advantages over other methods because it provides quantitative data and does not require instrumented fixators.

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Bone lengthening is a bone regeneration technique with multiple clinical applications. One of the most common complications of this treatment is the lack of adaptation of the surrounding soft tissue to their extension. A better understanding of the mechanobiology of the tissues involved in distraction osteogenesis would allow better control of the clinical cases. Bone lengthening treatments were performed in vivo in the metatarsus of Merino sheep, measuring the distraction forces by means of an instrumented fixator. The tissue relaxation after distraction was analyzed in this study. A viscoelastic model was also applied to distraction data to assess the mechanical behavior of the tissues during the distraction phase. Tissue relaxation is similar to other bone regeneration processes which do not imply surrounding soft tissue extension, e.g. bone transport. The effects of this tissue on distraction forces are limited to the first minutes of distraction and elongations above 4% of the original length with the protocol applied. Moreover, the surrounding soft tissue initially loses some of its viscoelasticity and subsequently suffers strain hardening from day 5 of distraction until the end of the distraction phase, day 15. Finally, anatomical changes were also evidenced in the elongated limb of our specimens.

Bone lengthening and bone transport are regeneration processes that commonly rely on distraction osteogenesis, a widely accepted surgical procedure to deal with numerous bony pathologies. Despite the extensive study in the literature of the influence of biomechanical factors, a lack of knowledge about their mechanobiological differences prevents a clinical particularization. Bone lengthening treatments were performed on sheep metatarsus by reproducing the surgical and biomechanical protocol of previous bone transport experiments. Several in vivo monitoring techniques were employed to build an exhaustive comparison: gait analysis, radiographic and CT assessment, force measures through the fixation, or mechanical characterization of the new tissue. A significant initial loss of the bearing capacity, quantified by the ground reaction forces and the limb contact time with the ground, is suffered by the bone lengthening specimens. The potential effects of this anomaly on the musculoskeletal force distribution and the evolution of the bone callus elastic modulus over time are also analyzed. Imaging techniques also seem to reveal lower bone volume in the bone lengthening callus than in the bone transport one, but an equivalent mineralization rate. The simultaneous quantification of biological and mechanical parameters provides valuable information for the daily clinical routine and numerical tools development.

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