Objectives and methods: Administration during root formation of a bisphosphonate, 1-hydroxyethylidene-1,1-bisphosphonate (HEBP), at a dose corresponding to 10 mg P/kg body weight, has been found to interfere with the formation of acellular cementum in rats. The purpose of this study was to measure the force required to extract a tooth lacking normal acellular cementum, and to correlate this force and the ultimate periodontal strength with the morphology of the periodontal tissues at different time intervals after single or multiple injections of HEBP.Results: A single injection of HEBP given during root formation inhibited the formation of acellular cementum and resulted in a temporary reduction of the extraction force and the ultimate tensile strength. Ninety days after the injection of HEBP, both parameters were the same as in the controls. The increase in extraction force and ultimate tensile strength was associated with the onset of occlusal contact of the teeth. The organization of periodontal ligaments was improved after the teeth reached the occlusal level. After daily injections of HEBP for 3 days, there was a permanent reduction in root length and dento-alveolar ankylosis developed in the furcation area.Conclusion: (1) A single or three injections of HEBP changed the formation of acellular cementum to that of an atypical hyperplastic cementum which increased the resorption risk at this site. (2) The ultimate tensile strength was markedly reduced in teeth lacking acellular cementum. (C) 1997 Elsevier Science Ltd.

The purpose of this study was to investigate the distribution of epithelial cells and the fate of the basement membrane along the root surface of rat molars during cementogenesis, and to test the hypothesis that the Hertwig's epithelial root sheath (HERS) cells remain on the root surface if mineralization is inhibited. To demonstrate the HERS cells and basement membrane, immunohistochemistry with antibodies against keratin and laminin were used. The dentin matrix mineralization was inhibited by a single injection of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP). A modified Gomori staining method was used to monitor the inhibition of mineral formation in dentin and cementum. Paraffin sections were stained with haematoxylin-eosin, and freeze-dried sections were used for Gomori and immunohistochemical stainings. We found that the formation of acellular cementum was suppressed above the dentin with inhibited mineralization. Instead, a hyperplastic matrix, different from acellular cementum, covered the dentin. This hyperplastic cementum had keratin- and laminin-positive cells incorporated; such cells were never incorporated in normal acellular cementum. The later formation of cellular cementum correlated, in controls, with the disappearance of HERS cells from the root surface. Treatment with HEBP resulted in a persistent presence of epithelial cells, interpreted as an inhibition of their disappearance. In conclusion, there is evidence that the cells of HERS are involved in the development of both acellular and cellular cementum. The developmental processes of these tissues appear in some way to be influenced by or associated with the initial mineralization of the dentin.

Cementum surface alterations induced by 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) in the maxillary first molars in rats were studied by means of scanning electron microscopy. Single or triple injections of HEBP inhibit the formation of acellular extrinsic fiber cementum and delay the formation of cellular mixed-fiber cementum. The results indicate the importance of acellular extrinsic fiber cementum as a protection barrier against root resorption and the different mechanisms underlying the formation of the two cementum varieties.

There are diverging opinions about the role of Hertwig's epithelial root sheath in the formation of cementum. In the present review this role is discussed. There is increasing evidence that Hertwig's epithelial root sheath is actively involved in the formation of both acellular and cellular cementum. The development of acellular cementum seems to be associated with secretion of enamel‐related proteins by cells of the epithelial root sheath. Formation of the matrix for cellular cementum appears to be induced by exposure of the inner layer of the epithelial root sheath to the mesenchymal cells in the dental follicle. Experimental studies with I‐hydroxyethylidene‐I, I‐bisphosphonate (HEBP) indicate that the formation of acellular cementum must be preceded by mineralization of the mantle dentin. If the mineralization is inhibited by means of HEBP, there is precocious separation of the two layers of the epithelial root sheath after which matrix for a cellular type of cementum is formed.

A single injection of 1-hydroxyethylidene-1, 1-bisphosphonate inhibits the formation of acellular cementum in rat molars. Instead, an atypical hyperplastic cementum is formed. In this study the resistance of this cementum to resorption was tested by applying an orthodontic force. It was found that roots lacking acellular cementum were readily resorbed. The readiness with which the atypical hyperplastic cementum was resorbed without any increase in multinucleated osteoblasts may offer useful opportunities to study the different phases of hard-tissue resorption.

In the developing tooth, the bisphosphonate HEBP causes hypoplasias and hypomineralization of the enamel and dentine as well as inhibition of acellular cementum formation. Here. we describe a novel effect, associated with dentine mineralization. HEBP was administrated to young rats, and the maxillary molars were analyzed histologically. Localized dentinal nodules, protruding towards the pulp, were found in the developing crown of the molars. They occurred in regions, where the mantle dentine was about to mineralize at the time of the injection, and were more frequent at the mesial cusp side. The nodules accumulated mineral, as evidenced by the fluorescence after calcein and tetracyclin labelling. Histologically, the nodules were separated from the enamel by a layer of mantle dentine and were progressively surrounded by predentine and dentine. The nodules were interpreted to contain transport or metabolism intermediates, which were locally accumulated due to the interruption of the mineralization process by HEBP.