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Technical Note

Validation of histologic bone analysis following Microfil vessel perfusion

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Pages 180-183 | Published online: 12 Nov 2013
 

Abstract

The ability to examine bone vascularity using micro-computed tomography following vessel perfusion with Microfil® and to subsequently perform histologic bone analysis in the same specimen would provide an efficient method by which the vascular and cellular environment of bone can be examined simultaneously. The purpose of this report is to determine if the administration of Microfil precludes accurate histologic assessment of bone quality via osteocyte count and empty lacunae count. Sprague–Dawley rats (n = 6) underwent perfusion with Microfil. Left hemi-mandibles were harvested, decalcified, and underwent vascular analysis via micro-computed tomography prior to sectioning and staining with Gomori’s trichrome. Quantitative histomorphometric evaluation was performed. Ninety-five percent confidence intervals (CIs) were used to determine statistical differences from an established set of controls (n = 12). Histologic analyses were successfully performed on specimens that had been perfused. Quantitative measures of bone cellularity of perfused versus control specimens revealed no statistical difference in osteocyte count per high-power field (95·33 versus 94·66; 95% CI: −7·64 to 6·30) or empty lacunae per high-power field (2·73 versus 1·89; 95% CI: −1·81 to 0·13). A statistical validation is reported that allows histologic analysis of cell counts in specimens which had been perfused with Microfil.

Funding was provided by National Institutes of Health grant RO1 CA 12587-01 to Dr Steven R. Buchman. The authors thank Megan Charms for her technical assistance with histomorphometric analyses.

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