Effects of Calcium and Phosphate on Secretion of Enamel Matrix and its Subsequent Mineralization in Vitro

¹ Laboratory of Tooth Development, Department of Oral Cell Biology, ACT A, Vrije Universiteit, de Boelelaan 1115, 1081 HV Amsterdam, The Netherlands

We examined the effects of various calcium (Ca) and phosphate (P) levels on enamel matrix synthesis, secretion, and its subsequent mineralization in vitro. Second maxillary molar tooth germs of three-day-old hamsters were cultured for nine days in vitro in media containing low ( 0.9 mmoI/ L), moderate ( 2.6 mmol/ L), or high (4.5 mmol/ L) medium levels of Ca, with either moderate ( 1.65 mmol/ L) or high ( 3.65 mmol/ L) medium levels of P. Explants were then examined histologically. For examination of matrix synthesis and mineralization, explants were labeled during the last 24 hr of culture with a triple label of 3H-proline, 45Ca, and 32PO4.

At low Ca levels, tooth germs failed to deposit enamel matrix and dentin, and no mineralization took place, regardless of the levels of P. Low levels of Ca, however, did not prevent deposition of pre-dentin. At moderate and high levels of Ca, considerable amounts of enamel and dentin were deposited in vitro, and both matrices mineralized. At high Ca levels, however, the forming enamel hypermineralized, was more irregular, and tended to be thinner. Increasing P concentrations at moderate and high Ca levels resulted in formation of a more regular enamel and dentin and a better-controlled mineralization. Biochemically, high levels of Ca tended to decrease enamel matrix secretion but significantly enhanced the uptake of 45Ca. This Ca-stimulated increase of 45Ca uptake could be reduced to below control levels by increases in P medium levels.

We conclude that: (1) a minimum medium Ca concentration is required to induce enamel matrix deposition and mineralization of both enamel and dentin; (2) high levels of medium Ca induce hypermineralization of enamel and give rise to deposition of a more irregular enamel than at moderate Ca levels; and (3) high levels of P are not able to induce mineralization when Ca levels are low but seem to moderate effects of moderate and high levels of Ca.