Methods to analyze bone regenerative response to different rhBMP-2 doses in rabbit craniofacial defects

Teja Guda, Aniq Darr, David T. Silliman, Maria H.R. Magno, Joseph C. Wenke, Joachim Kohn, Pamela R. Brown Baer

Research output: Contribution to journalArticlepeer-review


Multiple assessment methods are available to evaluate the performance of engineered scaffolds in accepted bone healing animal models. Evaluation and comparison of these methods can aid in the planning of future animal studies, as well as, inform clinical assessments as the engineered scaffolds translate into clinical studies and applications. To evaluate multiple bone assessment techniques, bone regrowth potential of tyrosine-derived polycarbonate (TyrPC) scaffolds loaded with various dosages of recombinant human bone morphogenetic protein-2 (rhBMP-2) (0, 10, 25, and 50μμg) was assessed after 16 weeks in vivo in a rabbit calvarial model. Traditional X-ray radiography and micro-computed tomography (micro-CT) analyses were used to quantify the volume and density of regenerated bone. Histomorphometric analysis was performed as the traditional gold standard of evaluation. While these techniques are fairly standard in bone tissue engineering, we also investigated 64-slice CT, a tool more commonly used clinically, for comparison and to guide translational efforts. The 64-slice CT scans were carried out at 4 and 16 weeks to monitor temporal bone healing patterns. Study results indicated a clear dose-dependent response of increasing regenerated bone volume with rhBMP-2 loaded on the TyrPC scaffolds after 16 weeks of implantation. Significantly more bone formation was observed at the highest dose of rhBMP-2 (50μμg), which is 25-50% of the previously recommended dose (100-200?μg) for this defect. A significant difference was observed between the lowest and highest doses using radiographs (p<0.001), micro-CT (p=0.002), and CT (p<0.001) and a high correlation was found between techniques (R2values between 0.446 and 0.911). It was found that the number of animals required per group to detect significant dose effects ranged between 6 and 8 for the imaging methods while histomorphometric analysis would require 25 animals per group to detect similar differences (desired power=0.9, α=0.05). Radiographic analysis provided quantifiable % defect coverage and radio-opacity, micro-CT provided spatial volumetric and bone density measures, histomorphometry provided biological confirmation, and 64-slice CT allowed for establishing of clinically relevant translational guidelines. These methodologies allow for a standardized and comprehensive description of bone regeneration and provide guidelines for the planning of future preclinical and clinical studies.

Original languageEnglish (US)
Pages (from-to)749-760
Number of pages12
JournalTissue Engineering - Part C: Methods
Issue number9
StatePublished - Sep 1 2014
Externally publishedYes

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering


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