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Limb Reconstruction with Grafts Derived from Decellularized Bone in a Growing Leporine Model
Elliot C Pennington, MD; Beatrice Dionigi, MD; Fabienne L Gray, MD; Azra Ahmed, BS; Joseph Brazzo III, BS; Andrey Dolinko, BS; Nathan Calderon, BS; Thomas Darrah, PhD; David Zurakowski, PhD; Ara Nazarian, PhD; Brian Snyder, MD, PhD; Dario O Fauza, MD, PhD
Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, MA

Background: We sought to examine a practical alternative for pediatric limb repair.

Methods: Growing New Zealand rabbits (n=12) with a complete, critical-size defect on the left tibiofibula were equally divided into two groups. One group received a decellularized, non-demineralized leporine tibiofibula graft. The other group received an equivalent graft seeded with mesenchymal stem cells labeled with green fluorescent protein (GFP), at a fixed density. Animals were euthanized at comparable time points 3-8 weeks post-implantation. Statistical analysis was by the Student t-test and Fisher’s exact test (p<0.05).

Results: There was no significant difference in the rate of non-union between the two groups, including on 3D micro-CT. Incorporated grafts achieved adequate axial bending rigidity, torsional rigidity, and both union yield and flexural strength, with no significant differences, or unequal variances between the groups. Correspondingly, there were no significant differences in extracellular calcium levels, or alkaline phosphatase activity. Histology confirmed the presence of neobone in both groups, with GFP-positive cells in the re-cellularized grafts.

Conclusion:s: Osseous grafts derived from decellularized, non-demineralized bone undergo adequate remodeling in vivo after repair of critical-size limb defects in a growing leporine model, irrespective of subsequent re-cellularization. This methodology may become a practical alternative for pediatric limb reconstruction.

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