The Effects of Synthetic Oligopeptide Derived from Enamel Matrix Derivative on Cell Proliferation and Osteoblastic Differentiation of Human Mesenchymal Stem Cells

Abstract

Enamel matrix derivative (EMD) is widely used in periodontal tissue regeneration therapy. However, because the bioactivity of EMD varies from batch to batch, and the use of a synthetic peptide could avoid use from an animal source, a completely synthetic peptide (SP) containing the active component of EMD would be useful. In this study an oligopeptide synthesized derived from EMD was evaluated for whether it contributes to periodontal tissue regeneration. We investigated the effects of the SP on cell proliferation and osteoblast differentiation of human mesenchymal stem cells (MSCs), which are involved in tissue regeneration. MSCs were treated with SP (0 to 1000 ng/mL), to determine the optimal concentration. We examined the effects of SP on cell proliferation and osteoblastic differentiation indicators such as alkaline phosphatase activity, the production of procollagen type 1 C-peptide and osteocalcin, and on mineralization. Additionally, we investigated the role of extracellular signal-related kinases (ERK) in cell proliferation and osteoblastic differentiation induced by SP. Our results suggest that SP promotes these processes in human MSCs, and that ERK inhibitors suppress these effects. In conclusion, SP promotes cell proliferation and osteoblastic differentiation of human MSCs, probably through the ERK pathway.

Amelogenin Exon 5 Peptide Promotes Cell Proliferation and Osteogenic Differentiation in Human Dental Pulp Stem Cells

“…In this study, we found that SP promoted the activity of ALP and the production of OCN. This result is similar to the effect of EMD and SP in human MSCs [13] and PDL stem cells [14]. Thus, we concluded that SP can also induce differentiation of DPSCs.…”

“…As well known, amelogenin can activate physiological actions through various signaling pathways. In our previous study, SP promotes the cell proliferation via the ERK pathway in human MSCs [13]. Therefore, we believe that SP promotes the cell proliferation via MAPK, such as ERK signaling.…”

“…Previous work has shown that amelogenin and EMD can promote the expression of all three genes in human osteoblasts and human dental pulp cells [28], as well as in human MSCs [29]. Similarly, SP can promote expression of ON in human PDL stem cells [13], and it promoted the expression of type I collagen in our previous in vivo study [11]. In this study, the SP enhanced the expression of Runx2, ON, and COL1A1 in DPSCs, suggesting that the SP enhances osteogenic differentiation in DPSCs during early osteogenic differentiation.…”

“…DPSCs at passage 3-4 were used for experiments. We used osteogenic medium containing 50 µM L-ascorbic acid 2-phosphate (Nacalai Tesque, Kyoto, Japan), 10 mM β-glycerophosphate (Wako Pure Chemical Industries Ltd., Tokyo, Japan), and 10 nM dexamethasone (Wako Pure Chemical Industries Ltd Tokyo, Japan), according previous our study [12][13][14].…”

“…Our previous study showed that SP induced bone and cementum tissue in rats [11]. We also showed that SP promotes cell viability in human periodontal ligament (PDL) fibroblasts [12], and promotes the osteogenic differentiation of mesenchymal stem cells (MSCs) [13] and PDL stem cells [14], as well as EMD.…”