A Reinvestigation of Murine Cranial Suture Biology: Microcomputed Tomography versus Histologic Technique

Abstract

Histology remains the standard form to analyze cranial suture in murine models, but this technique provides only limited "snapshots" of the entire suture and requires animal euthanasia with tissue destruction. Because of the bone complex microarchitecture, better methods are required to study the behavior of the cranial suture and its surrounding environment. The authors compared microcomputed tomography and histology as techniques to evaluate murine cranial sutures. A total of 360 microcomputed tomography images and 160 to 170 histologic sections were processed from a mouse at postnatal days 22 and 45, respectively. After euthanasia, the posterior frontal and sagittal sutures were imaged with a microcomputed tomography system and subsequently processed for histologic analysis. Quantitative analysis of two-dimensional images was performed to determine the percentage of bone in a 1-mm sample. Quantitative analysis of the percentage of bone within the sutures showed identical patterns by microcomputed tomography and histology techniques. Both methods demonstrated the posterior frontal suture to have heavier fusion patterns in the anterior and endocranial portions, with variable skip areas of complete patency on the endocranial surface, ectocranial surface, or both at day 45. Cranial suture fusion in the murine model is not an "all-or-none" phenomenon. The posterior frontal suture, previously thought to be completely fused on day 45 by histological analysis, showed variable fusion along the length of the suture by both methods. Quantitative assessment of the percentage of bone within the posterior frontal and sagittal sutures and morphologic assessment of these sutures demonstrated similar findings by both methods. Whereas thorough histologic evaluation of an entire suture would be extremely labor intensive and impractical, these findings help to validate microcomputed tomography as a rapid and reliable method of examining the entire suture in murine models.