Do different fluorochrome labels give equivalent histomorphometric information?

Substances that bind calcium are given to determine where and how fast bone is forming. Several vital dyes are used (tetracycline, calcein, alizarin, xylenol), but it is not known whether the histomorphometric results they provide are equivalent. This work tests whether different fluorochrome labels give the same results when they are quantitatively measured. Twelve-week-old rats (n = 58) were divided into six groups and given double labels IP of calcein, tetracycline HCl, alizarin complexone, or xylenol using a 1-7-1 scheme. Two other groups received either calcein followed by tetracycline, or tetracycline followed by calcein. Our results show that (a) tetracycline hydrochloride leads to a significant underestimation of mineralizing surface when given as the second label, probably because of its weak fluorescence; (b) there were no differences among any of the non-tetracycline labels for any of the histomorphometric measurements; (c) there was no evidence of osteoblast suppression with any label; and (d) there was no evidence that tetracycline caused increased osteoblast resting periods.