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Craniofacial Repair with Fetal Bone Grafts Engineered from Amniotic Mesenchymal Stem Cells
Fabienne L. Gray, MD, Christopher G. Turner, MD, Justin D. Klein, MD, Azra Ahmed, MS, David Zurakowski, PhD, Dario O. Fauza, MD
Department of Surgery, Children's Hospital Boston, Harvard Medical School, Boston, MA

Background: The ethically sensible realm of fetal tissue engineering can be expanded beyond life-threatening anomalies by amniotic fluid cell-based methods, as cell procurement poses no added maternal risk. We sought to determine whether osseous grafts engineered from amniotic mesenchymal stem cells (aMSCs) could be an alternative to craniofacial repair.
Methods: New Zealand rabbits (n=12) underwent creation of a full thickness diploic nasal bone defect. Animals were then equally divided in 2 groups based on how the defect was repaired, namely size-matched implants of electrospun poly (L-lactic) acid nanofibers with or without allogeneic aMSCs. Cell processing included phenotyping, nuclear labeling, nanoscaffold seeding at identical densities, and construct maintenance in osteogenic medium for 19-20 weeks. Animals were killed 8 weeks post-implantation for multiple analyses.
Results: Closure of the defect was noted in all subjects. Micro-CT scanning (2D and 3D) showed no significant differences in radiodensity between the groups. However, extracellular calcium levels were significantly higher in engineered grafts than in acellular implants (p=0.003), with no significant differences in alkaline phosphatase activity. Levene's test indicated significantly greater variability in mineralization in acellular implants than in engineered grafts by both direct calcium (P=0.008) and micro-CT measurements (p=0.032). Variances were not significantly different for alkaline phosphatase activity. Labeled cells were documented in the engineered grafts.
Conclusion: Craniofacial repair with osseous grafts engineered from amniotic mesenchymal stem cells lead to enhanced and more consistent mineralization when compared with an equivalent acellular prosthetic repair. Amniotic fluid-derived engineered bone may become a practical alternative for perinatal craniofacial reconstruction.


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