In vitro differentiation of epithelial cells cultured from human periodontal ligament

Background and Objective: Alkaline phosphatase and noncollagenous bone proteins are produced prior to cementum formation. While it has been suggested that epithelial rests of Malassez are involved in cementum formation, little is known about the relationship between epithelial rests of Malassez and cementum formation. The purpose of the present study was to determine whether the epithelial rests of Malassez cells cultured from human periodontal ligament can produce alkaline phosphatase and noncollagenous bone proteins, such as osteopontin, osteocalcin and bone sialoprotein. Material and Methods: An outgrowth of putative epithelial rests of Malassez cells was produced from periodontal ligament explant, and second passage cultures were used in the experiments. Human gingival epithelial cells and periodontal ligament fibroblasts were used as controls. The expression levels of amelogenin were analyzed by immunostaining and in situ hybridization. Furthermore, the expression levels of alkaline phosphatase and noncollagenous bone proteins were assessed by immunostaining and reverse transcription-polymerase chain reaction. Results: Amelogenin, alkaline phosphatase and osteopontin proteins and their corresponding mRNAs were detected at high levels in putative epithelial rests of Malassez cells. Osteocalcin and bone sialoprotein were not expressed in putative epithelial rests of Malassez cells. Alkaline phosphatase and noncollagenous bone proteins were seen in periodontal ligament fibroblasts, but not in gingival epithelial cells. Conclusion: Our results suggest that putative epithelial rests of Malassez cells cultured alone do not transform into maturing cells to form the cementum, but may play a potential role in the mineralization process.