Bone regeneration in cranial defects previously treated with radiation.
OBJECTIVE
Bone reconstruction in the head and neck region is frequently performed in the context of previous radiation treatment. Thus, the effectiveness of tissue engineering approaches for regenerating bone in radiated defects needs to be determined before considering application to patients. Incomplete healing is described when using osteoinductive protein therapy alone for bone defects previously treated with radiation. We hypothesized that a different approach using ex vivo gene therapy can heal these severely compromised defects.


METHODS
Animal study using Fisher rats.


METHODS
Two weeks before surgery, rats received either no radiation or a 12 Gray radiation dose to the calvarium. Syngeneic dermal fibroblasts were transduced ex vivo using an adenoviral vector containing the cDNA for bone morphogenetic protein (BMP)-7. Critical-sized calvarial defects were created, and either a transduced cell-seeded scaffold or an autologous bone graft was placed into the defect. Nonradiated defects were harvested 4 weeks later for both groups. Radiated defects treated with bone grafts were harvested at 4 weeks, and those treated with gene therapy were harvested either at 4 or 8 weeks. Gross inspection and histology were used to evaluate wound healing.


RESULTS
None of the bone grafts had gross or histologic evidence of healing at the wound margins. The nonradiated gene therapy treated defects revealed gross and histologic near-100% bone regeneration by 4 weeks after surgery. By gross inspection, the radiated defects had soft tissue admixed with islands of bone at both 4 and 8 weeks. The histologic appearance revealed areas of dense bone in a nonconfluent pattern admixed with adjacent cells having the morphologic appearance of hypertrophic chondrocytes, suggesting continued endochondral ossification.


CONCLUSIONS
Preoperative radiation significantly impairs the ability of BMP-7 ex vivo gene therapy to heal rat critical-sized cranial defects. This finding has significant implications for translating this tissue engineering approach to patients with cancer-related segmental bone defects.
Department of Otolaryngology--Head and Neck Surgery, Washington University School of Medicine, St. Louis, Missouri 63110, USA. nussenbaumb@ent.wustl.edu
Brian
B
Nussenbaum
R Bruce
RB
Rutherford
Paul H
PH
Krebsbach
D000818:Animals; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D001861:Bone Regeneration / Q000502:physiology*; D016025:Bone Transplantation; D003399:Craniotomy; D020405:Dermis / Q000473:pathology; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000473:pathology / Q000528:radiation effects / Q000637:transplantation; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006258:Head and Neck Neoplasms / Q000473:pathology / Q000532:radiotherapy; D019426:Integrases / Q000235:genetics; D011836:Radiation Tolerance; D011878:Radiotherapy / Q000009:adverse effects; D051381:Rats; D011916:Rats, Inbred F344; D012886:Skull / Q000473:pathology* / Q000528:radiation effects* / Q000601:surgery; D023822:Tissue Engineering / Q000379:methods; D014161:Transduction, Genetic; D016212:Transforming Growth Factor beta / Q000235:genetics; D014182:Transplantation, Autologous; D014764:Viral Proteins / Q000235:genetics
C525821:Bmp7 protein, rat; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D016212:Transforming Growth Factor beta; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases