MicroCT quantification of in vitro bone resorption of neonatal murine calvaria exposed to IL-1 or PTH

This study investigated how effectively a laboratory microCT (X-ray micro-computed tomography) system can quantify bone resorption in an in vitro calvarial model and how well this measure correlates with a conventional assay for calcium release (fluorometric titration). In vitro bone resorption in neonatal murine calvaria was quantified for 0.3 or 1.0 nM interleukin-1 (IL-1) or for 1.0 or 10.0 nM parathyroid hormone (PTH) treatment. Compared to control calvaria, a significantly greater fraction F of the calvarial "shell" (computed from the volumetric microCT data) was resorbed in treated calvaria of 5- to 7-day-old pups from the same litter. Excellent correlation (R2 = 0.8234) was observed between F and calcium release, and, unlike the calcium assay, the 3-D maps revealed where bone was resorbed. Mineral was preferentially lost near the sutures, and areas away from the suture were left relatively intact. MicroCT of calvaria before and after 96 h culture demonstrated that this X-irradiation neither increased control resorption nor prevented responses in the treated calvaria. Observations on calvaria from intact mice aged 1, 3, 5, 8, and 11 days showed uniformly distributed mineral (not a pronounced patchwork of "high" and "low" mineral regions) and increasing levels of mineral with age; this suggested that the spatial patterns of resorption were not related to inhomogeneities in the starting mineral distribution.