Alkaline Phosphatase Induces the Deposition of Calcified Layers in Relation to Dentin: An in vitro Study to Mimic the Formation of Afibrillar Acellular Cementum

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Abstract
An attempt was made to test the hypothesis that alkaline phosphatase, an enzyme which is abundant in periodontal ligament, plays a role in the formation of acellular root cementum. Thin slices of bovine dentin were incubated in Iscove Modified Dulbecco's Medium supplemented with 10% normal rabbit serum and 10 mmol/L (3-glycerophosphate (P-GP) or folded into pericardial explants. Intestinal bovine alkaline phosphatase (APase), covalently linked to agarose beads, was added to the cultures. In the presence of the enzyme, the dentin slices were covered with thin layers of mineralized material. Such layers were not observed in cultures not provided with APase-beads or β-GP. They also did not form in relation to demineralized dentin. The layers of calcified material appeared to consist of crystallites embedded in a granular matrix of moderate electron density, which often exhibited the presence of incremental lines and resembled the matrix of afibrillar acellular cementum formed under in vivo conditions. When pericardial explants were interposed between the enzyme-containing beads and the dentin, mineral deposition in relation to the dentin was retarded. This finding lends support to the view that soft connective tissues interfere with the free diffusion of phosphate.