Enamel Matrix Derivative Promote Primary Human Pulp Cell Differentiation and Mineralization

Abstract

Enamel matrix derivative (EMD) has been found to induce reactive dentin formation; however the molecular mechanisms involved are unclear. The effect of EMD (5–50 μg/mL) on primary human pulp cells were compared to untreated cells and cells incubated with 10−8 M dexamethasone (DEX) for 1, 2, 3, 7, and 14 days in culture. Expression analysis using Affymetrix microchips demonstrated that 10 μg/mL EMD regulated several hundred genes and stimulated the gene expression of proteins involved in mesenchymal proliferation and differentiation. Both EMD and DEX enhanced the expression of amelogenin (amel), and the dentinogenic markers dentin sialophosphoprotein (DSSP) and dentin matrix acidic phosphoprotein 1 (DMP1), as well as the osteogenic markers osteocalcin (OC, BGLAP) and collagen type 1 (COL1A1). Whereas, only EMD had effect on alkaline phosphatase (ALP) mRNA expression, the stimulatory effect were verified by enhanced secretion of OC and COL1A from EMD treated cells, and increased ALP activity in cell culture medium after EMD treatment. Increased levels of interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemoattractant proteins (MCP-1) in the cell culture medium were also found. Consequently, the suggested effect of EMD is to promote differentiation of pulp cells and increases the potential for pulpal mineralization to favor reactive dentine formation.

Potential Role of Long Non‐Coding RNA in Osteogenic Differentiation of Human Periodontal Ligament Stem Cells

“…In the present study, target genes chosen for CNC analysis were reported to be closely related to osteogenic differentiation. [20][21][22][23][24][25][26] Notably, lncRNA AC078851.1, upregulated 26-fold after osteoinduction, was found to be negatively correlated with IL6; and lncRNA TCONS_00007046, upregulated 14.80-fold after osteoinduction, was found to be positively correlated with ALP and BMP2. The consistency of expression level and correlational relationship suggests that these lncRNAs may be involved in the osteogenesis process by influencing related mRNAs and are worth further study.…”

“…mRNAs reported to be related to osteogenic differentiation were chosen as targets, including: 1) ALP; 2) BMP6; 3) COL1A1; 4) COL1A2; 5) IL6; 6) BMP5; and 7) BMP2. [20][21][22][23][24][25][26][27] The CNC network included 131 negative pairs and 262 positive pairs, indicating that mRNAs and lncRNAs were closely correlated. One mRNA was correlated with one or more lncRNAs and vice versa (Fig.…”

“…Except for BMP2, RUNX2, and BMP4, the following are also accepted to be crucial for osteogenesis: 1) BMP5; 2) alkaline phosphatase (ALP); 3) collagen Type I (COL1A); 4) msh homeobox (MSX) 1; and 5) interleukin (IL) 6. [21][22][23][24][25][26][27] The functional mechanism of these osteogenesis-related mRNAs has been discussed in depth, but their connection with lncRNAs is little studied and needs to be further investigated.…”

“…Remarkably, Zhuang et al 20 found that upregulation of lncRNA maternally expressed ( MEG ) 3 promoted osteogenic differentiation of MSCs from patients with multiple myeloma by altering transcription of BMP4 . Except for BMP2, RUNX2 , and BMP4 , the following are also accepted to be crucial for osteogenesis: 1) BMP5 ; 2) alkaline phosphatase ( ALP ); 3) collagen Type I ( COL1A ); 4) msh homeobox ( MSX ) 1; and 5) interleukin ( IL ) 6 21–27 . The functional mechanism of these osteogenesis‐related mRNAs has been discussed in depth, but their connection with lncRNAs is little studied and needs to be further investigated.…”

Gene expression of collagen type I alpha 2 and its relationship with dental fluorosis

“…9 In dental fluorosis the activity of ameloblasts, osteoblasts, osteoclasts, osteocytes, matrix formation and calcium homeostasis are altered.…”