Degradative Changes in Whole Enamel Homogenates IncubatedIn Vitroin the Presence of Low Calcium Ion Concentrations

The purpose of this study was to investigate overall degradative changes occurring to enamel matrix proteins in small, freeze-dried pieces of rat incisor enamel homogenized and incubated directly for 0-48 hours in a synthetic enamel fluid solution (165 mM total ionic strength with 0.153 mM calcium chloride) versus other samples homogenized and incubated for the same time intervals in distilled water. The results indicated that many alterations in the apparent molecular weights of enamel matrix proteins took place under both conditions although the rates for many degradative changes over a 48 hour period were often slower in distilled water than in synthetic enamel fluid. Freeze-dried enamel samples homogenized and incubated in 165 mM Tris-HCl buffer at pH 8.0 showed changes comparable to those seen with distilled water. This suggested that differences observed between samples incubated in enamel fluid versus distilled water were unrelated to pH or ionic strength of the solutions and may be the result of a requirement by some enamel proteinases for small amounts of free calcium ions in incubation media. Of interest were findings that some enamel matrix proteins, especially those in strips taken from the first half of the secretory stage of amelogenesis, were degraded much faster in distilled water than in synthetic enamel fluid. The reasons for this effect are unclear although, in this case, calcium ions could be inhibitory to hydrolysis of certain matrix proteins by the enamel proteinases.