Tetracycline labeling of bone in vivo

We have analyzed various aspects of tetracycline labeling technique for the measurement of bone apposition rate in vivo. Our efforts were restricted to those aspects that are frequently questioned when data obtained using this technique are interpreted as representing the rate of bone apposition. Rat bone was labeled in vivo by sequential injections of oxytetracycline at a dose range of 3 to 24 mg/kg body weight and at intervals ranging from 24 to 72 h. The bone apposition rate was calculated by measuring the distance from the first dose of label to the subsequent ones. As these distances are by far too small to be determined accurately by any available micrometer eyepiece, we have used a scanning microscope photometer which allows measurements on slow-forming sites that otherwise would have been considered nongrowing sites. Using these techniques, we have demonstrated that oxytetracycline has no effect on the bone apposition rate when used in the concentrations indicated. In addition, we found that at labeling intervals of 96 h or more, periods of osteoblastic inactivity are likely to be included in measurements at individual sites. The instantaneous apposition rate is thus underestimated at these long time intervals.