Quantitative histomorphometric assessment of regenerate cellularity and bone quality in mandibular distraction osteogenesis after radiation therapy

Abstract

Background: The use of mandibular distraction osteogenesis (MDO) for tissue replacement after oncologic resection in head and neck cancer could have immense therapeutic ramifications. We have previously demonstrated significantly decreased mechanical and microdensitomeric metrics of our MDO regenerate after 36-Gy radiation. Quantitative histomorphometry, a third metric, would permit objective investigation of the effects of radiation on tissue and cellular composition. Our hypothesis is that radiation-induced cellular depletion and diminution in function impair optimal bone regeneration.

Methods: Five rats received radiation to the left mandible; 5 received none. All animals underwent surgical placement of external fixators, creation of mandibular osteotomies, distraction to a 5.1-mm gap width, and consolidation. Point counting and color thresholding were performed.

Results: There was a significant increase in empty lacunae and a corresponding diminution in osteocytes after radiation. Whereas the volume fraction of mineralized, mature bone was not different, that of nonmineralized, immature osteoid was significantly increased in the radiated group compared with that in the nonradiated group.

Conclusions: Our findings confirm our prior 2 metrics. Actually, all 3 diverse metrics--microdensitometry, biomechanical analysis, and histomorphometry--corroborate our hypothesis of cellular depletion and diminution of function as the potential mechanism of radiation-induced attenuation in the distracted regenerate. Furthermore, our findings of tissue and cellular changes in the irradiated regenerate elucidate the pathophysiology of decreased bone quality when amalgamated with our previous results. Therapeutic agents may now be introduced, and their effects on the irradiated regenerate critically measured, so that MDO may be used as a viable reconstructive option in patients with head and neck cancer.

FIGURE 1

A, Group 1 MDOx timeline. B, Group 2 MDO timeline.

FIGURE 2

Rat undergoing radiation. Window in lead shield permits radiation only to left posterior hemimandible.

FIGURE 3

Rat’s left-sided mandibular osteotomy being distracted, no anesthesia required.

FIGURE 4

A, Sagittal section of midportion of rat mandible stained with Gomori trichome. Green trapezoid outlines distraction regenerate. Bone is highlighted in purple by color thresholding. B, Sagittal section of midportion of rat mandible stained with Gomori trichome. Green trapezoid outlines distraction regenerate. Nonmineralized matrix is highlighted in red by color thresholding.

FIGURE 5

A single digital photograph at 16× magnification demonstrating bone stained green with Gomori trichome, osteocytes within lacunae, and EL.

FIGURE 6

Side-by-side comparison of single digital photos at 16× magnification, of MDO and MDOx rats.

FIGURE 7

A, Graph of mean Oc/HPF in MDOx and MDO. B, Graph of mean EL count per HPF in MDOx and MDO. C, Graph of mean values of nonmineralized matrix/OV per total volume in MDOx and MDO.