Abstract
Amelogenin (AMG) is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG (rhAMG) on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts (OCCM-30) were treated with different concentrations (0.1, 1, 10, 100, 1000, 10,000, 100,000 ng · mL-1) of recombinant human AMG (rhAMG) and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen type I (COL I), osteopontin (OPN), runt-related transcription factor 2 (Runx2), cementum attachment protein (CAP), and alkaline phosphatase (ALP) genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer. Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomal-associated membrane protein-1 (LAMP-1), which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100,000 ng · mL-1 increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG (100,000 ng · mL-1) upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100,000 ng · mL-1 remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10,000 and 100,000 ng · mL-1 rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue.
Highlights
Enamel matrix derivative (EMD) is a group of enamel matrix proteins that are derived from Hertwig’s root sheath of developing porcine teeth
The OCCM-30 cells used in this study are immortalized
There were no significant differences among recombinant human AMG (rhAMG) concentra- mouse cementoblasts that are well characterized and widely tions until 72 h, when an apparent increase was noted in the used for cementoblast stimulation in vitro to investigate 100,000 ng · mL-1 group (Fig. 1)
Summary
Enamel matrix derivative (EMD) is a group of enamel matrix proteins that are derived from Hertwig’s root sheath of developing porcine teeth. In vitro studies have shown positive effects of EMD on the proliferation of periodontal ligament (PDL) cells, gingival fibroblasts, follicle cells, and cementoblasts.[1,2,3,4,5,6,7] Commercially available EMD suspended in a hydrogel (Emdogain®, Institut Straumann AG, Basel, Switzerland) is used for regenerative therapy around natural teeth. This formulation is a specific approach for enhancing periodontal regeneration. It is known that EMD can interact with various other cells, including osteoblasts and dental stem cells.[4,18,19,20,21,22,23]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
