The use of the stable isotope 44Ca in studies of calcium incorporation into dentin

The incorporation into rat incisor dentin of two calcium isotopes, the stable ⁴⁴Ca and the radioactive ⁴⁵Ca, was studied using secondary ion mass spectrometry (SIMS) stepscanning and imaging, and autoradiography, respectively. The results demonstrated a time-dependent incorporation of the calcium isotopes into the mineral phase of dentin. With the SIMS step-scanning, detecting ⁴⁴Ca, the ion yield was high in the odontoblasts 2 min after intravenous injection. After 10 min a marked increase in signal intensity was found at the dentin mineralization front. This result was consistent with those obtained by ⁴⁵Ca autoradiography; a peak of incorporation occurred 10 min after injection of the isotope. Likewise, localization of ⁴⁴Ca to the mineralization front could be demonstrated 10 min after injection by SIMS imaging. In images obtained at earlier intervals, no such increase in ion yield could be detected. The results show that the nonradioactive, stable isotope ⁴⁴Ca can be used as a marker for biomineralization in a similar way to radioactive ⁴⁵Ca.