J Periodontal Implant Sci.  2011 Feb;41(1):30-43. 10.5051/jpis.2011.41.1.30.

Analysis of gene expression during mineralization of cultured human periodontal ligament cells

• Choi HD
• Noh WC
• Park JW
• Lee JM
• Suh JY

• KMID: 2212133
• DOI: http://doi.org/10.5051/jpis.2011.41.1.30

Abstract

PURPOSE
Under different culture conditions, periodontal ligament (PDL) stem cells are capable of differentiating into cementoblast-like cells, adipocytes, and collagen-forming cells. Several previous studies reported that because of the stem cells in the PDL, the PDL have a regenerative capacity which, when appropriately triggered, participates in restoring connective tissues and mineralized tissues. Therefore, this study analyzed the genes involved in mineralization during differentiation of human PDL (hPDL) cells, and searched for candidate genes possibly associated with the mineralization of hPDL cells.
METHODS
To analyze the gene expression pattern of hPDL cells during differentiation, the hPDL cells were cultured in two conditions, with or without osteogenic cocktails (beta-glycerophosphate, ascorbic acid and dexamethasone), and a DNA microarray analysis of the cells cultured on days 7 and 14 was performed. Reverse transcription-polymerase chain reaction was performed to validate the DNA microarray data.
RESULTS
The up-regulated genes on day 7 by hPDL cells cultured in osteogenic medium were thought to be associated with calcium/iron/metal ion binding or homeostasis (PDE1A, HFE and PCDH9) and cell viability (PCDH9), and the down-regulated genes were thought to be associated with proliferation (PHGDH and PSAT1). Also, the up-regulated genes on day 14 by hPDL cells cultured in osteogenic medium were thought to be associated with apoptosis, angiogenesis (ANGPTL4 and FOXO1A), and adipogenesis (ANGPTL4 and SEC14L2), and the down-regulated genes were thought to be associated with cell migration (SLC16A4).
CONCLUSIONS
This study suggests that when appropriately triggered, the stem cells in the hPDL differentiate into osteoblasts/cementoblasts, and the genes related to calcium binding (PDE1A and PCDH9), which were strongly expressed at the stage of matrix maturation, may be associated with differentiation of the hPDL cells into osteoblasts/cementoblasts.

Keyword

Microarray analysis; Gene expression profiling; Periodontal ligament; Cell differentiation

MeSH Terms

Adipocytes
Adipogenesis
Apoptosis
Ascorbic Acid
Calcium
Cell Differentiation
Cell Movement
Cell Survival
Connective Tissue
Durapatite
Gene Expression
Gene Expression Profiling
Homeostasis
Humans
Microarray Analysis
Oligonucleotide Array Sequence Analysis
Periodontal Ligament
Stem Cells
Ascorbic Acid
Calcium
Durapatite

Figure

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  Figure 1 Alizarin red S staining of the mineralized nodules cultured for 7 days (A) and 14 days (B) with osteogenic medium and non-osteogenic medium. Circular photographs show human periodontal ligament cells in the culture plate (24 wells). On day 7, mineralized nodules were not observed at all, but on day 14, mineralized nodules were only observed in the osteogenic group.

• 

  Figure 2 Expression on day 7 and 14 of selected genes by human periodontal ligament (hPDL) cells cultured in osteogenic or non-osteogeneic medium. The hPDL cells were cultured for 7 days in an osteogenic medium (+) or non-osteogenic medium (-). The 12 out of 14 genes were comparable with DNA microarray data. The expression patterns of tachykinin, precursor 1 were not significant, and nebulette were not expressed. Similar results were obtained in 2 separate experiments and representative data are shown. GAPDH: glyceraldehyde-3-phosphate dehydrogenase, PDE1A: phosphodiesterase 1A, calmodulin-dependent, APOL4: apolipoprotein L, 4, MTP18: mitochondrial protein 18 kDa, CYTL1: cytokine-like 1, PHGDH: phosphoglycerate dehydrogenase, PSAT1: phosphoserine aminotransferase 1, IGSF4: cell adhesion molecule 1, IL6: interleukin 6 (interferon, beta 2), NEBL: nebulette, FBN2: fibrillin 2, TAC1: tachykinin, precursor 1, CXCL12: chemokine (C-X-C motif ) ligand 12, SLC16A4: solute carrier family 16, member 4 (monocarboxylic acid transporter 5), ALDH1A3: aldehyde dehydrogenase 1 family, member A3.

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