Introduction: The long consolidation phase of patients undergoing distraction osteogenesis (DO) causes a high risk of side effects and contributes to high costs. Thus, the development and evaluation of treatments that accelerate the bone consolidation process is of great interest. Evidence suggests that recombinant human bone morphogenetic protein 2 (rhBMP-2) increases the mechanical integrity of the callus. However, the potential benefits of rhBMP-2 on trabecular microarchitecture during DO have not been investigated up to date. In this study the regenerate microarchitecture was assessed using 3D micro-computed tomography (CT).

Methods: Mid-diaphyseal osteotomies were created in the right limb of twenty-four skeletally mature sheep, which were stabilized with an external fixator. After a latency period of 4 days, the tibiae were distracted at a rate of 1.25 mm daily over a period of 20 days. The operated limbs were randomly assigned to three treatment groups and one control group: (A) triple injection of rhBMP-2/NaCl, (B) single injection of rhBMP-2/Hydroxylapatite, and (C) single injection of buffer/Hydroxylapatit, (D) no injection. Groups A and C were injected at day 27. Group B was injected on days 3, 10 and 17. The animals were sacrificed after 74 days. The tibiae were analyzed by CT and for bone volume/total volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th.), trabecular separation (Tb.Sp.) and Connectivity. The BV/TV was maesured for the total volume of the distraction zone (BV/TVtotal) respectively in a subvolume with emphasize on the cortical bone region (BV/TVcortical). All other microarchitecture parameters were measured in the cortical weighted subvolume.

Results: The stereologic evaluation revealed a significant higher BV/TVcortical, Tb.N and Connectivity in the triple rhBMP-2 injected group A than in the control (D). Furthermore, the Tb.Sp. in group A was significant lower than in group D. The single injections of rhBMP-2/carrier in group B showed a significant higher BV/TVcortical, Tb.N and Connectivity than the control (D). Although the BV/TVcortical was increased in group A and B, there was no significant difference in BV/TV total between the rhBMP-2 treated groups (A, B) and the control (D).

Discussion: In this DO model a triple injection of rhBMP-2 has been demonstrated to induce significant changes in trabecular microarchitecture. RhBMP-2 does not increase the total amount of newly formed bone, but it enhances the formation of the corticalis. The microstructural changes in the cortical volume: increase of Tb.N and Connectivity, decrease of Tb.Sp., are discussed to be biomechanically highly relevant. This study suggests that rhBMP-2 optimizes the trabecular microarchitecture, which might explain the advanced mechanical integrity of newly formed bone under rhBMP-2 treatment.