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Alkaline phosphatase (ALP) is a glycoprotein thought to be involved in processes leading to mineral formation in tissues like bone and cementum. In the rat molar periodontium, several regions are associated with the formation of cementum (periodontal ligament, inner part of the gingiva), whereas other areas are not (e.g., the outer part of the gingiva just beneath the outer oral epithelium). In an attempt to establish how the spatial distribution of ALP activity relates to cementum formation, we assessed the activity of the enzyme quantitatively in the periodontium of the rat maxillary molars, by using the indoxyl-tetrazolium salt method. It appeared that the distribution of enzyme activity in the ligament was heterogeneous, indicating local variations in the phosphate household. Highest activity was found in areas related to mineralization, adjacent to the alveolar bone and cementum. Enzyme activity was higher adjacent to cellular cementum than to acellular cementum. With respect to acellular cementum, a highly significant positive correlation was found between ALP activity and cementum thickness, which indicates a close relationship between local phosphate production and cementum formation rate. An interesting observation in the connective tissue of the gingiva mesial to the first molar was a sharp demarcation between an ALP-positive inner part, adjacent to the tooth, and an ALP-negative outer part, underneath the outer oral epithelium. In the interdental gingiva, the entire connective tissue proved positive for the enzyme, suggesting that this region consists of the combined inner gingival parts of two adjacent teeth.

Fibroblasts of the periodontal ligament, by their alkaline phosphatase (ALP) activity, are considered to play a role in the formation of acellular cementum. As a means of exploring this hypothesis, periodontal ligament explants from rat incisors were cultured in direct contact with bovine dentin slices in the presence of 10 mmol/L beta-glycerophosphate. Periosteal and pericardial tissue explants were maintained under similar conditions. After two weeks, the slices were harvested and processed for electron microscopic examination. Controls included periodontal ligament explants to which the ALP-inhibitor levamisole was added. The results suggest that only ALP-positive cultures from periodontal ligament and periosteum form mineralized layers along the dentin. After demineralization, layers consisted of fine filamentous or granular material of moderate electron-density and resembled afibrillar acellular cementum. Our findings support the hypothesis that periodontal ligament fibroblasts, by means of their ALP activity, play a pivotal role in the formation of acellular cementum.

The spatial distribution of alkaline phosphatase (ALP) activity was examined in the periodontal ligament of the continuously growing rat incisor. With the indoxyl-tetrazolium salt method, enzyme activity was demonstrated in undecalcified cryosections, and the amount of reaction product was quantified. ALP activity appeared to be distributed heterogeneously. Its highest activity was found in the bone-related compartment of the ligament. In the tooth-related compartment and the supracrestal extension of the ligament, enzyme activity was significantly lower, but still higher than in the lamina propria of the gingiva. In the part of the ligament bordering the cementum, highest activity was found in the apical region just occlusal to Hertwig's epithelial root sheath, where formation of acellular cementum begins. From there toward the incisal edge, the activity of the enzyme gradually decreased. It is suggested that differences among the various parts of the periodontal ligament are related to local variations in phosphate metabolism and cementum deposition.