Differential Effects of Amelogenin on Mineralization of Cementoblasts and Periodontal Ligament Cells

Abstract

Amelogenin is a major component of developing extracellular enamel matrix proteins and plays a crucial role during the formation of tooth enamel. In addition, amelogenins are suggested to exert biologic functions as signaling molecules through cell-surface receptors. The purpose of this study is to examine the effect of recombinant human full-length amelogenin (rh174) on the mineralization of human cementoblasts (HCEMs) and human periodontal ligament cells (HPDLs). HCEMs, namely, a cell line immortalized by transfection of human telomerase reverse transcription gene, and HPDLs isolated from human first premolars were cultured and treated with 0 to 1,000 ng/mL rh174. The messenger ribonucleic acid (mRNA) levels of alkaline phosphatase (ALP), osteocalcin (OCN), and bone sialoprotein (BSP) were examined by real-time polymerase chain reaction analysis. The protein levels of OCN and BSP were examined by Western blot analysis. ALP activity and calcium deposition of cell cultures were also determined. Mineralization of cells was evaluated by red dye staining. The treatment of HCEMs with rh174 upregulated the ALP, OCN, and BSP mRNA levels. In addition, the protein levels of OCN and BSP, ALP activity, and calcium deposition were enhanced, resulting in enhanced mineralization. Conversely, there were no significant effects of rh174 on the mineralization of HPDLs. The present study shows that rh174 enhances mineralization accompanied by upregulation of mineralization markers in HCEMs, whereas it has no effect on that in HPDLs, suggesting different effects of amelogenin on PDL and cementum.