The Enhancing Effects of Amelogenin Exon 5-Encoded Peptide from Enamel Matrix Derivative on Odontoblast-Like KN-3 Cells

Hirohito Kato,Kazuya Tominaga,Makoto Umeda,Yoichiro Taguchi,Muneyasu Shida,Yu-Wei Tsai,Yi-Chie Chen,Kazutaka Imai,Reiko Taguchi,Yaru Ruan

doi:10.3390/app8101890

Abstract

Enamel matrix derivative (EMD) is applied for periodontal therapy. We created a synthetic amelogenin peptide (SP) derived from EMD, and have previously investigated the biological function of SP. However, it is unknown whether SP affects odontoblastic differentiation. In this study, we tested the effects of SP in the odontoblast-like cells, KN-3 cells. KN-3 cells were cultured with SP (0 to 1000 ng/mL) and then cultured for 3, 8, 24, or 48 h in order to determine the effects of SP on cell proliferation and detect its optimum concentration. KN-3 cells were treated with SP in odontogenic differentiation medium cultured for 3 or 7 days. Odontogenic markers were measured by the detection of alkaline phosphatase (ALP) activity and dentin sialo phosphoprotein (DSPP) expression, the calcified nodule formation, and calcium deposition. The addition of SP significantly promoted cell proliferation at 100 ng/mL, generating the greatest change in cell proliferation. SP also showed increased odontogenic expression markers and mineralization. These results suggest that SP, derived from EMD, could have potential for application in dental pulp capping.

Highlights

Enamel matrix derivative (EMD) can induce the formation of hard tissue, such as alveolar bone and cementum tissue [1,2]
We found that 100 ng/mL of synthetic peptide (SP) significantly promoted KN-3 proliferation at 8, 24, 48, and 72 h (Figure 1A, p < 0.05)
We found that SP enhanced alkaline phosphatase (ALP) activity, which is similar to the effect of EMD in hard tissues and the effect of SP in KN-3 cells

Summary

Introduction

Enamel matrix derivative (EMD) can induce the formation of hard tissue, such as alveolar bone and cementum tissue [1,2]. EMD is applied for periodontal therapy and bone regeneration. We previously showed that subcutaneous injections of EMD can induce the growth of cartilage tissue and eosinophilic round bodies (ERBs) [3]. We further analyzed these ERBs by using MALDI-TOF, and found fragments of exon 5 of amelogenin. We found that the SP could induce bone-like tissue formation in artificial periodontal defects in rats [5,6]. We found that SP could enhance the cell proliferation of periodontal ligament (PDL) cells [7] and enhance osteoblastic

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Discussion

Conclusion