Muneyasu Shida

The Effects of Amelogenin Exon 5 Encoded Peptide from Enamel Matrix Derivative Enhances in Odontoblast like Cells, KN-3 Cells

Enamel matrix derivative (EMD) is used for periodontal tissue regeneration therapy. We 21 designed a synthetic amelogenin peptide (SP) derived from EMD, and have previously investigated 22 the biological function of SP. However, it is unknown whether SP affects odontoblastic 23 differentiation. In the present study, we investigated the effects of SP in odontoblast-like cells, KN24 3 cells. KN-3 cells were treated with SP (0, 1, 10, 100, or 1000 ng/mL) and then cultured for 3, 8, 24, 25 or 48 hours, in order to determine the effects of SP on cell proliferation and detect its optimum 26 concentration. To investigate the effect of SP on odontogenic differentiation, KN-3 cells were treated 27 with SP in odontogenic differentiation medium cultured for 3 or 7 days. Odontogenic differentiation 28 was performed by measuring alkaline phosphatase (ALP) activity, the mRNA expression of dentin 29 sialophosphoprotein (DSPP), the formation of calcified nodules, and calcium deposition into the 30 extracellular matrix. The addition of SP significantly promoted KN-3 cell proliferation; a 31 concentration of 100 ng/ml generated the greatest change in cell proliferation. SP also showed 32 increased expression of markers of odontogenic differentiation and mineralization. These results 33 suggest that SP, derived from EMD, could be a potential for applicate to the dental pulp capping. 34